The optimal time for blood testing is in the range from 1 to 4 hours after collection. In the range from 5 to 30 minutes, platelets temporarily adapt to the anticoagulant and their aggregation occurs, which can lead to a false decrease in them in the blood sample.

It is not advisable to examine blood later than 8 hours after collection, because Some cell characteristics change: the volume of leukocytes decreases, the volume of red blood cells increases, which ultimately leads to erroneous measurement results and incorrect interpretation of the results. Only the hemoglobin concentration and platelet count remain stable during 24 hours of blood storage.

Blood should not be frozen. Capillary blood with K 2 EDTA should be stored at room temperature and analyzed within 4 hours after collection.

To ensure high-quality research results, it is necessary to clearly control the time and storage conditions of samples before performing analysis.

If a delayed analysis is necessary (transportation over long distances, technical malfunction of the device, etc.), blood samples are stored in the refrigerator (4 o - 8 o C) and examined within 24 hours. However, it should be taken into account that cell swelling and changes in parameters associated with their volume occur. In practically healthy people, these changes are not critical and do not affect quantitative parameters, but in the presence of pathological cells, the latter can change or even be destroyed within several hours from the moment the blood is taken.

Immediately before the test, the blood should be thoroughly mixed for several minutes to dilute the anticoagulant and uniformly distribute the formed elements in the plasma. Long-term constant mixing of samples until they are examined is not recommended due to possible injury and decay of pathological cells.

Blood testing on an automatic analyzer is carried out at room temperature. Blood stored in the refrigerator must first be warmed to room temperature, since at low temperatures the viscosity of the blood increases and the formed elements tend to stick together, which in turn leads to impaired mixing and incomplete lysis. Examination of cold blood may cause “alarm signals” due to compression of the leukocyte histogram.

When performing hematological studies at a considerable distance from the place of blood collection, problems associated with unfavorable transportation conditions inevitably arise. The impact of mechanical factors (shaking, vibration, stirring, etc.), temperature conditions, the likelihood of spillage and contamination of samples can affect the quality of analyzes. To eliminate these reasons, when transporting blood tubes, it is recommended to use hermetically sealed plastic tubes and special insulated transport containers. During transportation, contact of the sample (with native material) and the referral form is not allowed, in accordance with biological safety rules.

^ 4.3. Reception and registration of biological material

Tubes with venous blood samples are delivered to the laboratory on the day of collection in racks in special bags for the delivery of biological material, in which the tubes must be in a vertical position, and when transported over a remote distance - in special containers.

The laboratory employee receiving the material must check:

Correctness of registration of the referral: the referral form indicates the data of the subject (last name, first name and patronymic, age, medical history or outpatient card no., department, diagnosis, therapy performed);

Labeling of tubes with blood samples (they must be marked with a code or surname of the patient, identical to the code and surname in the form for sending material for research). The laboratory assistant must register the delivered material and note the number of tubes.

^ 4.4 Calculation of the cellular composition of blood on a hematology analyzer

Calculation of the cellular composition of blood on a hematology analyzer must be carried out in strict accordance with the instructions for the device.

When working on a hematology analyzer you should:

Use blood stabilized with K-2 EDTA in the recommended proportion for a specific salt and disposable tubes for hematological studies (simple or vacuum);

Before analysis, carefully but thoroughly mix the test tube with blood (it is recommended for this purpose to use a device for mixing blood samples - a rotomix);

Use reagents registered in accordance with the established procedure; when changing reagents to products from another manufacturer, you should check and establish the calibration of the device;

Start the device, observing all stages of washing, achieving zero (background) values ​​of indicators in all channels;

Pay attention to device messages about possible systematic errors: remember that an increase in MSHC above normal values ​​is usually the result of a measurement error;

If an error is detected, be sure to eliminate the cause and do not work on a faulty device;

Perform quality control procedures according to the appropriate program. with an assessment of the result obtained;

Work meaningfully, comparing the results obtained with the clinical characteristics of the patient’s samples;

After completing measurements, thoroughly rinse the analyzer;

When stopping the device for a long period of time, be sure to carry out all preservation procedures (if possible, together with a service engineer);

If technical problems arise, you should seek the help of a service engineer from a company that has a license for technical maintenance; Do not trust the work to untrained personnel.

When starting to work on a hematology analyzer, you should take into account the limits of linearity of measurement of the analyzed parameters. Evaluating samples with analyzed parameter values ​​exceeding the measurement linearity limit may lead to erroneous results. In most cases, when analyzing samples with hypercytoses, the analyzer displays a “D” (dilute) icon instead of the value of the measured parameter, which indicates the need to dilute the sample and repeat the measurement. Dilutions must be carried out until similar final results are obtained with the two nearest dilutions.

^ EXAMPLE ─ Analysis of a sample from a patient with hyperleukocytosis on three different hematological analyzers is presented in the table.

Table 3

¦The results of counting the amount for hyperleukocytosis, performed on three different hematological analyzers

^ Hematology analyzers	Sample dilution degree				Total value
	^ Whole blood	1:1	1:3	1:4	WBC
1	D	274,5	274,5	143,1	715,5
2	322,6	253,2	177,7	141,8	709,0
3	D	D	168,3	139,1	695,5

The table shows that when analyzing whole blood, only one analyzer (2nd) gave a digital value for the number of leukocytes, the other two indicated the need to dilute the sample. Subsequent step-by-step measurement of the sample in three dilutions made it possible to achieve similar final results on all instruments only with dilutions of 1:3 and 1:4. Thus, the final leukocyte count was 700.0 - 710.0 x 10 9 /l, which is more than 2 times higher than the initial value obtained in whole blood on the 2nd analyzer and 1.5 at a 1:1 dilution on the 1st and 2nd analyzers.

1. 
   ^ Calibration of hematology analyzers

Calibration refers to the procedure for setting up the analyzer, resulting in

Which achieves the equality of the systematic component of the error to zero (zero offset) or confirms that the offset is equal to zero taking into account the calibration error. Calibration of hematology analyzers can be performed in two main ways:

For whole blood;

Based on stabilized blood samples with certified values.

^ 5.1 Calibration using whole blood

Whole blood calibration can be performed by comparison with a reference analyzer, the calibration of which has been verified and can be taken as reference.

For calibration by comparison with a reference analyzer, 20 patient venous blood samples randomly selected from the daily research program of a laboratory with a reference analyzer are used. Blood should be collected in vacuum tubes with anticoagulant K 2 EDTA. After analysis on a reference analyzer, the blood tubes are delivered to the laboratory for analysis on a calibrated device. The time between measurements on the reference and calibrated analyzer should not exceed 4 hours. Analysis of calibration samples on the instrument being calibrated should be performed in the same manner as analysis of patient samples. The results obtained are entered into a table in the form given in Appendix 1. These results are used to determine the corresponding calibration factors, which are numerically equal to the slope coefficients of the calibration lines passing through the origin of coordinates on the graph.

Calibration factors are calculated on a computer using the EXCEL program, which is part of the Microsoft OFFICE package of any version. To calculate calibration factors in this program, a scatter diagram is constructed, where measured values ​​are plotted along the X axis, reference values ​​are plotted along the Y axis, and a regression line is set through the origin of coordinates. The program draws a calibration line and determines its slope, which is the desired calibration factor. If gross errors are detected - points significantly removed from the calibration line on the graph, they must be removed from the calculation (erasing the entire line in the EXCEL table). The resulting calibration factor is entered into the table in Appendix 1 and is used to manually adjust the calibration of the device in accordance with its operating instructions.

1. 
   ^ Calibration using stabilized blood samples with certified values

This calibration method is based on commercial calibration and control materials, which are an artificial mixture of stabilized human red blood cells and fixed human or animal cells that mimic leukocytes and platelets. Due to the significant difference in the biological properties of the components of these materials from natural blood, the certified values ​​of hematological parameters depend on the specific type of analyzer.

Whenever possible, calibrators that are specifically designed by the manufacturer to calibrate a particular type of analyzer should be used. The set point errors in modern hematology calibrators are generally consistent with medical requirements. However, such calibrators are not available for all types of analyzers. In addition, due to the limited shelf life (usually no more than 45 days), it is not always possible to obtain samples that are not expired.

Control materials are not intended by the manufacturer for calibration purposes: the tolerances for values ​​are much wider, and the certification and production control methods are not as strict as for calibrators. When calibrating, control materials can be used only as one source of information about the metrological properties of the device.

It should, however, be borne in mind that when using both control materials and calibrators for calibration purposes, the features of sample preparation and operation of the analyzers have a significant impact on the components of the systematic error. These influencing factors cannot be taken into account when calibrating with commercial materials. As a result, in all cases, the calibration performed must be verified by analyzing the statistics of the results obtained in patient samples (erythrocyte indices, normal ranges).

5.2.1 Calibration frequency of hematology analyzers

Requirements for calibration frequency are usually contained in the analyzer's operating instructions. Typically, calibration is required after repair or when significant drift is detected in internal laboratory quality control procedures.

5.2.2 Calibration procedure

The calibration procedure is carried out in accordance with the operating instructions for the analyzer.

5.2.3 Monitoring the calibration result

This control consists of analyzing the calibration sample immediately after calibration. The completed calibration is accepted if the analysis results correspond to the calibration values, taking into account the analytical variation of the analyzer.

5.2.4 Calibration verification using red blood cell indices

This method of checking and refining calibration takes advantage of the fact that the average values ​​of red blood cell indices - MCHC, MCH and MCV across patients have quite a small variation and can therefore be effectively used for calibration control and further quality control. The recommended number of samples for averaging is 20. Any patient samples can be used to determine the mean MCHC, while patients with anemia should be excluded to determine the mean MCHC and MCV.

Many modern hematology analyzers have built-in programs for calculating current averages, which greatly facilitates data processing.

Target values ​​of erythrocyte indices when averaged over 20 samples of patients aged 18 to 60 years, regardless of gender, are shown in Table 4.

Table 4

Target values ​​of erythrocyte indices

If the average values ​​obtained are outside the control tolerance, this means that the MCV or Hgb or RBC calibration needs to be adjusted.

When determining the reasons for unsatisfactory calibration of red blood cell parameters when using commercial calibration/control materials, the following assessment of the reliability of the certified values ​​of the materials should be taken into account:

The most accurate and time-stable parameter is the concentration of red blood cells;

A stable parameter is Hgb, but it may depend on the reagents used (for example, cyanogen or cyanogen-free);

The most unstable parameter is MCV. The MCV value tends to change over the entire shelf life of the material over a fairly wide range of values. When refining MCV calibration, data obtained from the analysis of average red blood cell indices take precedence over certified values ​​in stabilized blood samples.

Assuming that the RBC calibration is correct, Table 5 shows the possible reasons for the average index values ​​to fall outside the control tolerance.

Table 5

Reasons for going beyond the average values ​​of erythrocyte indices

Erythrocytic indices	Average index value	Average index value	Average index value	Average index value
MCHC	Below the border	Above the border	Above the border	Below the border
MCH	Within tolerance limits	Within tolerance limits	Above the border	Below the border
MCV	Above the border	Below the border	Within tolerance limits	Within tolerance limits
Cause	MCV is too high	MCV lowered	Hgb is too high	Hgb is low

Instructions for collection, storage and transportation

biological material for laboratory research

Biological material must be in labeled tubes (containers) and accompanied by a completed individual referral form in one copy. It is necessary to carefully and legibly fill out the referral, indicating the following information: date, test tube number, name of the healthcare facility that sent the biomaterial for testing, name of the doctor who collected the material; Full name, age and gender of the patient, preliminary diagnosis; during pregnancy - gestational age. Reception of biological material: PCR and immunochemical blood tests- With 09:00 before 18:00 except weekends and holidays. Bacteriological studies- With 09:00 before 11:00 except Fridays, weekends, pre-holidays and public holidays.

for identifying pathogens using PCR method

1.1.1. The biomaterial is taken from the suspected habitat of microorganisms and the development of infection.

1.1.2. The amount of material collected should be small. Excess discharge, mucus and pus negatively affect the quality of DNA extraction and contribute to DNA degradation during storage and transportation.

1.1.3. When introducing biomaterial taken from a patient with a probe with a cotton swab or brush into an Eppendorf tube with a buffer solution, it is necessary:

Maintain sterility;

Before immersing the biomaterial collected on the swab (brush) into the solution, smear it on the dry wall of the test tube, then moisten the swab (brush) in the solution and, rotating the probe, thoroughly rinse off all the material from the wall of the test tube and the swab (brush);

If possible, squeeze the swab against the wall of the test tube, remove the probe with the swab (brush) and close the test tube.

1.2. Methods for collecting biomaterial for PCR research

1.2.1. Scrapings. For scrapings, disposable sterile probes with a cotton swab with increased adsorption or “brushes” are used, less often - Volkmann spoons, small ear spoons or similar instruments with slightly blunted edges. Using scraping movements, collect the material. The material under study should contain the largest possible number of epithelial cells and a minimum amount of mucus, blood and exudate. Add the material to a sterile Eppendorf tube with buffer solution (see section 1.1.).

Scraping from the cervical canal: Before taking a scraping, it is necessary to remove excess mucus with a sterile cotton swab and treat the cervix with sterile saline. Insert the probe into the cervical canal to a depth of 0.5-1.5 cm, avoiding contact with the walls of the vagina, and collect the material with a scraping movement (not until there is blood). A small number of red blood cells in the sample does not affect the analysis result. In the presence of cervical erosion, material is taken from the border of healthy and altered tissue.

Urethral scraping: insert the probe into the urethra (in men - to a depth of 2-4 cm) and collect the material with several rotational movements. The day before taking the material, provocation (spicy food, alcohol, etc.) is allowed. It is recommended to refrain from urinating for 1-2 hours before taking the sample. If there is heavy purulent discharge, scraping should be taken no later than 15 minutes after urination.

Conjunctival scraping taken after anesthetizing the eye with a 0.5% dicaine solution. Having everted the eyelid, use a probe with a cotton swab (or an eye scalpel) to collect epithelial cells from the conjunctiva.

Rectal scraping: Insert the probe into the anus to a depth of 3-4 cm and collect the material with a rotational movement.

1.2.2. Strokes. Using a disposable sterile probe with a cotton swab or brush, take a small amount of discharge (from the vaginal vault, pharynx, nasopharynx, etc.) and transfer it to a sterile Eppendorf tube with a buffer solution (see section 1.1.).

1.2.3. Blood. For PCR studies, blood must be native(not curled).

Collect blood aseptically, by venipuncture, into a proprietary test tube (2-3 ml) with EDTA anticoagulant ( Heparin cannot be used as an anticoagulant!), mix gently, gently turning the test tube.

Blood with EDTA is a biomaterial for the following groups of studies:

- genetic research(the moment of blood sampling does not matter).

- identification of infectious agents(the most informative are samples taken during chills, fever, i.e., presumably during viremia or bacteremia).

1.2.4. Urine. Collect the first or mid-morning urine in an amount of at least 10 ml in an empty sterile container with a tightly screwed lid. If urine collection is carried out during the day, then before this the patient should not urinate for 1.5-3 hours.

1.2.5. Sputum. In the morning, sanitize the mouth and throat (rinse with a solution of baking soda). Collect sputum in an empty, sterile, wide-mouth bottle.

1.2.6. Biopsy. The material is taken from the area where the infectious agent is supposed to be located, from damaged tissue or the area bordering the damage. Place the biopsy sample in a sterile Eppendorf tube with buffer solution.

1.2.7. Saliva, gastric juice, cerebrospinal fluid, synovial fluid. Saliva (1-5 ml), gastric juice (1-5 ml), cerebrospinal fluid (1-1.5 ml) - place in an empty sterile tube (container).

1.2.8. Prostate juice. After massage of the prostate gland, the juice is collected in an amount of 0.5-1 ml into an empty sterile tube (container). If it is impossible to obtain juice, immediately after the massage, collect the first portion of urine in an amount of no more than 10 ml (this portion contains prostate juice).

1.2.9. Washings, bronchoalveolar lavage (BAL). A sterile cotton swab and 5-7 ml of physiological solution are used to rinse, for example, from the tip of an endoscope or bronchoscope, and place 0.5-5 ml of the rinse into an empty sterile tube (container).

1.2.10. Feces. A probe with a cotton swab is placed inside the stool and turned slightly, capturing a small amount of material. The material is then placed in a sterile Eppendorf tube containing a buffer solution.

1.3. Rules for storing biological material for PCR research

1.3.1. Smears, scrapings:

In the refrigerator (+4ºС - +8ºС) for up to 7 days;

In the freezer (-20ºС) for up to 14 days (only one-time defrosting is allowed!).

1.3.2. Urine: in the refrigerator (+4ºС - +8ºС) no more than 1 day;

1.3.3. Blood with EDTA: in the refrigerator (+4ºС - +8ºС) - to determine infectious agents - no more than 1 day; for genetic research – up to 4 days. Cannot be frozen.

1.3.4. Sputum

1.3.5. Biopsies:

In the refrigerator compartment (+4ºС - +8ºС) no more than 1 day,

In the freezer (-20ºС) for up to 2 weeks.

1.3.6. Prostate juice: in the refrigerator (+4ºС - +8ºС) for no more than 1 day.

1.3.7. Synovial fluid: in the refrigerator (+4ºС - +8ºС) for no more than 1 day.

2. Rules for collecting, storing and transporting blood

for immunochemical studies

2.1.1. On the eve of blood collection, avoid physical activity, stressful situations, physiotherapeutic procedures, taking medications (except in cases of medication prescribed by a doctor), oral contraceptives, drinking alcoholic beverages and fatty foods. Do not smoke immediately before the test.

2.1.2. When studying the function of the thyroid gland during treatment with drugs containing thyroid hormones, the study is carried out 24 hours after the last dose of the drug; 2-3 days before taking blood, avoid taking medications containing iodine.

2.1.3. When studying PSA a week before the test, exclude any manipulation of the prostate gland.

2.1.4. Blood is taken in the morning, on an empty stomach (to avoid chylosis, i.e. turbidity of the serum), under conditions of physiological rest. Before blood collection, the patient must be given a 15-minute rest.

2.1.5. Blood is taken in the treatment room of a healthcare facility, with the patient in a “sitting” or “lying down” position, from the ulnar vein in compliance with the rules of asepsis and antisepsis. Blood is collected into a sterile tube, monovet syringe or blood collection system (Vacutainer®, Vacuette®).

2.1.6. To obtain serum, venous blood is allowed to settle until it clots completely. After the formation of a fibrin clot, the latter is separated from the walls with a sterile glass rod, strictly individual for each sample, the tube is centrifuged at 3000 rpm for 15 minutes * .

* Monovet syringes and blood collection systems (Vacutainer®, Vacuette®) are centrifuged without prior manipulation.

2.1.7. The yellow supernatant (serum) is carefully pipetted into an empty capped tube (5 ml).

2.2. Rules for storing blood and serum for immunochemical studies

2.2.1. TOlevel: in the refrigerator (+40C - +80C) for 1 day.

2.2.2.Serumblood:

In the refrigerator at a temperature of +40C - + 80C for 4 days;

In the freezer at -200C for 2 weeks.

Only one defrosting is allowed!

ATTENTION! The subject of examination is serum that does not contain admixtures of red blood cells, bacterial growths, chyle, or hemolysis. If any of these signs are present, the serum is destroyed and a repeat blood draw is prescribed.

2.3. Rules for collecting and storing blood for lupus anticoagulant testing

And coagulation studies

2.3.1. Blood is taken in the morning on an empty stomach from the ulnar vein, observing the rules of asepsis and antisepsis on the day of the study. To prevent activation of blood clotting, vein compression should not exceed 1 minute .

2.3.2. If the study is prescribed while taking medications that affect blood clotting, this must be noted in the referral.

2.3.3. The blood is collected in a special tube containing sodium citrate, to the mark. Failure to comply with this rule changes the blood/anticoagulant ratio, which may adversely affect the accuracy of the result.

2.3.4. Within 1 hour after drawing blood, the tube must be centrifuged at 3000 rpm for 15 minutes.

2.3.5. Plasma storage: up to 6 hours at a temperature of +40C - +80C or up to 2 weeks at a temperature of -200C.

2.4. Rules for collecting and storing blood for the study of glycated (glycosylated) hemoglobin

2.4.1. Blood is taken in the morning on an empty stomach from the ulnar vein, observing the rules of asepsis and antiseptics.

2.4.2. The blood is collected into a special tube containing EDTA and mixed smoothly. Do not centrifuge!

2.4.3. Delivery to the Laboratory on the day of blood collection.

2.4.4. Storage: up to 2 weeks at -200C.

2.5. Rules for prenatal screening

2.5.1. Optimal timing of the study: I ​​trimester – 10-13 weeks of pregnancy; II trimester – 16-18 weeks of pregnancy.

2.5.2. When sending for a study, a special direction is filled out, in which it is necessary to indicate the individual data of the pregnant woman: age (day/month/year), weight, ultrasound results (CTR, BPR, number of fetuses, gestational age according to ultrasound (weeks + days); if available - data on the size of the cervical fold - NT) with the obligatory indication of the date of the ultrasound and the name of the doctor who performed the ultrasound; the presence of additional risk factors (smoking, diabetes, IVF), ethnicity.

2.5.3. To screen for the second trimester of pregnancy, you can use ultrasound data performed in the first trimester.

3. Rules for taking, storing and transporting biomaterial

for bacteriological research

3.1. The biomaterial is taken before applying a course of antibacterial therapy or no less than a week after its completion.

3.2. To prevent contamination of the sample by microorganisms from the external environment, the taken biomaterial is transferred into an empty container or test tube with a transport medium in compliance with standard sterility rules: the stopper of the test tube (container) is opened/closed so that the inside of the stopper remains sterile; sterile containers do not stay open for long.

3.3. Taking biomaterial for identification mycoplasmas and ureaplasmas, trichomonas, gonococci, nonspecific microflora, cultures of anaerobic bacteria produced in branded tubes with carbon transport medium:

Open the package in the indicated place according to the diagram;

Remove the test tube and open it, discard the lid;

Use a probe to take a scraping/smear, a biopsy sample, or dip the probe into a biological fluid;

Immediately place the probe with the biomaterial into the test tube (the probe handle is the lid of the test tube);

Number the test tube and indicate the number in the direction.

3.4. Types of biological material for taking into the transport coal environment: smears/scrapings, prostate juice, cerebrospinal fluid, synovial fluid, biopsy.

3.5. Storage of biomaterial in tubes with transport medium for detection mycoplasmas and ureaplasmas, nonspecific microflora until the moment of transportation -

Delivery to the Laboratory within one day.

3.6. Storage of biomaterial in tubes with transport medium for detection Trichomonas, gonococci, cultures of anaerobic bacteria until the moment of transportation - at room temperature.

3.7. Native biomaterial: sputum, prostate juice, synovial fluid from 1 to 10 ml for the detection of nonspecific microflora, mycoplasmas and ureaplasmas are collected in an empty sterile container.

It is necessary to notify the Laboratory the day before or on the day of taking the biomaterial before 11:00.

Storage of native biomaterial until transportation - in the refrigerator compartment (+4ºС - +8ºС).

Delivery to the Laboratory on the day of collection of the material.

3.8. Urine for testing the degree of bacteriuria(average morning portion, 10-20 ml) is collected in an empty sterile container with a lid after thorough toileting of the external genitalia.

It is necessary to notify the Laboratory the day before or on the day of taking the biomaterial before 11:00.

Storage of biomaterial until transportation - in the refrigerator compartment (+4ºС - +8ºС).

Delivery to the Laboratory on the day of collection of the biomaterial.

3.9. Blood for sterility.

The most informative is a blood sample taken during a rise in temperature.

Blood is collected in branded bottles with a two-phase transport medium (bottle for adults, bottle for children). Carefully open the plastic cap of the bottle and wipe the emerging part of the rubber stopper with 70% alcohol. Under sterile conditions, take blood from a vein with a syringe and inject it through a rubber stopper into the bottle (4 ml into a bottle for adults, 2 ml into a bottle for children).

It is necessary to notify the Laboratory the day before or on the day of taking the biomaterial before 11:00.

Storing blood until transport - at room temperature.

Delivery to the Laboratory on the day of collection of the biomaterial.

3.10. Breast milk to identify nonspecific microflora, it is taken from each breast into a separate sterile hermetically sealed cup. Label the cups: “left breast”, “right breast”.

Before collecting the biomaterial, wash the breasts with warm water and soap, wipe with a clean towel, carefully treat the nipples and the nipple area with a cotton swab moistened with a 70% solution of ethyl alcohol, and dry with a sterile cloth (each gland is treated with a separate swab). The first 10-15 ml of expressed milk is not used for analysis. Express the second portion of milk from each breast into a labeled cup in an amount of ~ 10 ml.

It is necessary to notify the Laboratory the day before or on the day of taking the biomaterial before 11:00.

Delivery to the Laboratory within 2 hours on the day the material is taken.

3.11. Feces for dysbacteriosis.

Before taking a sample, drinking alcohol for 3 days and taking antibiotics for 2 weeks is prohibited. Stool must be obtained without enemas or laxatives. For children with constipation, a bar of soap can be used for an irritating effect. Feces are collected in an individual vessel, washed from disinfectant. Using a sterile spoon, take 3-5 g of an average portion of feces and place it in a sterile container.

It is necessary to notify the Laboratory the day before or on the day of taking the biomaterial before 11:00.

Storage until transportation - store the container with biomaterial wrapped in cotton wool and paper to preserve heat.

Delivery to the Laboratory on the day of collection of the biomaterial.

4. Rules for taking, storing and transporting biomaterial

for microscopic studies

4.1. From each patient - 2 glasses(one glass - spare). On every glass - 3 points: vagina ( V), cervical canal ( WITH), urethra ( U). Apply strokes so that the matte surface is on the left, on top, and from it - from left to right: v, c, u.

4.2. The marking (No.) on the glass should not dissolve in alcohol (you can sign on the frosted part of the glass with a simple pencil). Smears should be fixed by air drying (30-60 min).

4.3. The glasses must be marked, folded side by side, packed in an individual bag with the appropriate accompanying inscription (health care facility, patient's full name, number on the glass).

4.4. The package and completed referral are sent to the Laboratory.

5. Rules for taking, storing and transporting biomaterial

5.1. For 24 hours before the study, douching, intravaginal therapy, and sexual intercourse should be avoided. You cannot take material during menstruation. The most informative are samples taken 14-20 days after the start of menstruation.

5.2. The material is taken before bimanual examination, various diagnostic tests, and before taking the material for PCR. The material is taken without preliminary treatment of the vaginal mucosa.

5.3. A scraping from the vaginal/cervical mucosa is taken with a nylon Cervex-Brush, spatula or other instrument and spread evenly over the glass, leaving one end of the glass clean (for marking). One point is taken per glass and duplicated onto another spare glass (total of two glasses per point). Smears should be fixed by air drying (30-60 min).

5.4. The glasses must be marked (No. and full name of the patient), folded side by side, packed in an individual bag with the appropriate accompanying inscription (health care facility, full name of the patient, No. on the glass).

5.5. Dried smears can be stored for up to 6 days at room temperature.

5.6. The package and completed directions (each point has its own direction indicating the type of biomaterial) are sent to the Laboratory.

Do not put the direction in a bag with glasses, do not wrap the glasses in the direction!

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"Nizhny Novgorod Medical College"

(GBOU SPO NO "NMK")

Test

Topic: “Organization of reception, labeling and registration of biomaterial”

Completed:

student of group 221-IV lab.

Stepanychev M.N.

Teacher:

Belova M.N.

N. Novgorod 2016

Introduction

1. Theoretical part

1.1 Registration of directions

1.2 Laboratory research modes

1.3 Conditions for storage and transportation of material for clinical laboratory research

1.4 Scope of application

1.5 Organization of the process of transporting biomaterial by courier and the responsibility of the courier

1.6 Reception of biomaterial in the laboratory

1.7 Transfer of biomaterial and referrals to the relevant departments of the laboratory

1.8 Registration of order forms by operators

2. Practical part

Conclusion

Introduction

Laboratory tests are the most sensitive indicators of the patient's condition. Clinical laboratory diagnostics specialists have long realized that many factors can affect the results of laboratory tests. Among these factors: correctly delivered and competently designed samples of biomaterial and accompanying documentation. In the absence of such supporting information (factors that influence laboratory tests), the clinician may misinterpret the result and take the wrong action for the patient.

The purpose of this test is to draw attention to the need to revise the rules for receiving and processing documentation of biological material. Proper application of information should lead to a reduction in unnecessary research, reduced costs, and a better understanding of research results.

Target:

Study the organization of reception, labeling and registration of biomaterial.

Tasks:

1. Study of the use of biomaterial.

2. Preparation of its documentation.

1. Theoretical part

Currently, laboratory research methods play a key role in the diagnosis and treatment of diseases. When examining patients, the organizational aspects of laboratory diagnostics, ensuring the accuracy and correctness of laboratory tests, the use of stable, technically reliable analytical methods, and quality control of research are of great importance.

Healthcare institutions use a fairly wide range of laboratory methods, and along with unified routine techniques, modern research methods that are highly specific and informative are also used.

The introduction of highly specific laboratory methods for diagnosing diseases into healthcare practice requires standardization at all stages of the diagnostic process.

The share of errors at the preanalytical stage in the total number of laboratory errors ranges from 50% to 95%. Errors that occur at the preanalytical stage of laboratory research devalue the entire further course of laboratory research, lead to the loss of significant funds, and discredit laboratory methods in the eyes of the attending physician due to the unreliability of the results obtained.

For all types of laboratory research, the preanalytical stage combines a set of processes before the actual laboratory analysis, which consists of:

· thorough preparation of the subjects;

· obtaining biomaterial (blood, urine, etc.);

· transport of biomaterial to the laboratory;

· primary processing of biomaterial in the laboratory;

· distribution and storage of biomaterial in the laboratory.

The main task of these stages of the preanalytical process is to ensure the stability (safety) of the components of biomaterials taken for research and to minimize the influence of various factors that change their quality.

In this regard, the preanalytical stage is considered.

1.1 Registration of directions

When taking material, you must fill out the directions correctly. Test tubes and accompanying documents and labels must not be mixed up. The compartment and serial number are indicated on the tube. On the test tube with blood to determine group and Rh affiliation - additionally the patient's last name.

In the direction for analysis when taking biomaterial, you must indicate: (all data taking into account the order of the Ministry of Health of the Republic of Belarus dated September 28, 2007 No. 787 on primary medical documentation forms)

· patient data - last name, first name, patronymic, date of birth, gender, address, medical history number, diagnosis.

· the name of the doctor who sent the biomaterial for analysis;

date and time of taking the biomaterial;

· give a brief description of the biomaterial being sent (indicate the type of biomaterial); material that does not correspond to the declared one will not be examined - for example, saliva instead of sputum, serum instead of plasma;

· features of collection of biomaterial (place of collection - for blood, for urine - volume, portion, time of collection);

· all necessary studies, and not just a “biochemical blood test”.

The referral for analysis is signed by the doctor.

Avoid duplication of tests.

1.2 Laboratory research modes

All blood should be supplied only in centrifuge, chemically clean tubes taken in the laboratory. Blood for research in emergency analysis mode should be delivered separately and not placed on the table together with scheduled tests without warning. The department employee who delivered the urgent test must ensure that the test is accepted by the laboratory employee.

Emergency tests are performed within 2 hours, only for patients in serious condition, if emergency intervention is necessary. To perform urgent analysis, a separate laboratory assistant and a laboratory diagnostics doctor are assigned. Carrying out research in urgent analysis mode requires greater consumption of reagents and specialist working time, and as a result, greater financial costs.

Currently, the urgent analysis laboratory performs the following blood tests on an emergency basis:

Determination of hemoglobin concentration

Hematocrit calculation

· Counting the number of red blood cells

· Leukocyte count

· Leukocyte count

Platelet count

On-call analysis is performed during the working day - for newly admitted patients or when the patient’s condition worsens and does not require emergency intervention.

Note: Normal laboratory values ​​refer to fasting samples. Therefore, the interpretation of the results of emergency and routine research cannot consist of a simple comparison with normal values.

Planned studies are all studies scheduled the day before, and the material for which arrives at the laboratory from 7 to 9 am. Execution time is up to 6 hours; in some cases, a longer analytical procedure is required.

Laboratory examination of employees is carried out at the direction of the doctor responsible for medical examination of employees. In all other cases, examination is provided on a paid basis.

1.3 Conditions for storage and transportation of material for clinical laboratory research

In order to prevent occupational infection, any biological fluid must be considered as a potentially infectious material, observing all relevant rules for handling it during transportation, storage and processing. Biomaterials from infectious patients must have special labeling and must be handled with extreme caution. All biomaterials from patients with HIV infection must be marked in the direction “code 120” with the obligatory indication of the medical history number.

The resulting biological fluid must be delivered to the laboratory as quickly as possible. If it is necessary to determine glucose and acid-base status, blood should be delivered to the laboratory immediately.

At all stages of transportation and processing, blood should be kept in tubes, closed with lids, to prevent evaporation and contamination by microbes and various external substances.

Upon delivery, the tubes should be positioned vertically, with the lids up, which helps to preserve the samples and accelerate the formation of a clot when receiving serum, reduces shaking during transportation and the risk of hemolysis.

Test tubes should not be filled to the brim. Whole blood obtained without anticoagulants should not be refrigerated prior to delivery to the laboratory to avoid hemolysis.

Centrifugation is carried out no later than 1 hour after taking the biomaterial.

1.4 Scope of application

This standard operating procedure (hereinafter referred to as SOP) defines the procedure for receiving, registering biological material entering the laboratory from the procedure room of (name of institution) and other institutions served by the laboratory, as well as identifying inconsistencies and eliminating them.

This SOP is intended for the laboratory assistant and laboratory operator.

1.5 Organization of the process of transporting biomaterial by courier and the responsibility of the courier

The temperature in the refrigerator, which is indicated on the display inside the car, is recorded by the courier in the journal “Logbook of temperature conditions in car refrigerators”, and in this journal the courier indicates his full name, registration plate of the car and the time of departure of the car for biomaterial immediately before traveling to the institutions .

The courier, having arrived at the health care facility, goes to the registry of this institution, where he picks up containers with various biomaterials. Each container contains the name of the institution and the type of biomaterial (blood, urine, feces, slides with smears, scrapings). The courier places the containers horizontally in the refrigerator of the machine, as shown in the photo.

The courier is responsible for the integrity of the containers, their safety, for ensuring the proper temperature conditions in the refrigerator of the car (+4 - +8 o C), as well as for the safety of the biomaterial delivered from institutions to the laboratory. During the journey, the courier monitors the temperature in the car’s refrigerator, which is displayed on a display installed in the car’s interior.

1.6 Reception of biomaterial in the laboratory

The temperature indicated on the display in the refrigerator of the car at the time of removal of the containers is noted by the courier in the journal “Logbook of temperature conditions in car refrigerators” in the column corresponding to the state number of the car. And also in the same log, the driver indicates the time of arrival of the car with biomaterial from institutions.

The courier hands over labeled containers with blood samples, smears and scrapings to the medical laboratory assistant.

In the office, a medical laboratory assistant opens the lid of the container and removes tubes with blood, slides with smears and scrapings, and folders with directions for research.

Sorts blood tubes separately into racks, according to the type of tubes (biochemical, hematological and coagulology) and the names of the institutions that are indicated on the racks.

1.7 Transfer of biomaterial and referrals to the relevant departments of the laboratory

In the office, a medical assistant laboratory assistant places racks with test tubes for biochemical, immunological, coagulological studies in containers marked “for transferring biomaterial” and takes them for centrifugation.

The paramedic-laboratorian calls the hematology and PCR departments so that the paramedic-laboratory assistants from the relevant departments take the biomaterial.

Next, he transfers referrals for research to registration operators.

1.8 Registration of order forms by operators

Registration procedure:

- the operator reads the barcode with a scanner, pasted on the direction form;

- then the operator enters the patient’s passport data into the LIS: full name, date of birth, residential address and other data: source of order (compulsory medical insurance, voluntary health insurance, cash payment, clinical examination), institution number, department, full name of the doctor who ordered the examination, diagnosis, MES code ( medical and economic standard).

- after this, the operator enters into the LIS those indicators that the attending physician prescribed and saves the generated order in the LIS.

2. Practical part

City Clinical Hospital No. 40 was opened in 1966 and currently represents the only multidisciplinary medical complex in Nizhny Novgorod, which includes: a hospital, a maternity hospital, an antenatal clinic, a children's and adult clinic. It is the main clinical base of the Department of Surgery, a center for advanced training and professional retraining of specialists, the Nizhny Novgorod State Medical Academy, which has been operating on the basis of the hospital since its opening. A functional feature of Hospital No. 40 is the reception of patients with almost any pathology in emergency cases around the clock. Ambulances deliver more than 100 patients daily.

The hospital and its structural divisions have created all the necessary conditions to provide timely assistance to patients and high-quality treatment. Traditional methods are confidently used and improved here, and the latest medical technologies are actively introduced. Modern diagnostic equipment is being used everywhere. Over the past two years, as part of the healthcare modernization program, the hospital has received 216 units of medical equipment, including modern endoscopic, laboratory, and ultrasound diagnostic equipment, which made it possible to perform more complex interventions in hepatopancreatology, coloproctology, and phlebology.

Leading specialists take an active part in specialized seminars, conferences, and symposia at the Russian and international level. laboratory clinical biomaterial

City Hospital No. 40 is a wide-ranging clinical base where modern scientific developments and techniques are applied in practice. Here, joint work has been carried out for a long time and fruitfully with the Department of Surgery of the Center for Advanced Training and Professional Retraining of Specialists of the Nizhny Novgorod State Medical Academy. Its organizer, Professor Igor Leonidovich Rotkov, became the founder of the surgical school, which almost all the hospital’s surgeons attended and today continue to develop their accumulated experience, introducing new developments and techniques. A city phlebological center has been created on the basis of the hospital. For several decades, the Department of Pediatric Surgery has had a scientific department of pediatric gastroenterology as a branch of the Nizhny Novgorod Institute of Pediatrics. On the basis of the maternity hospital there are departments of obstetrics and gynecology for advanced training of doctors and training of students of the NSMA. Here specialists from many regions of Russia improve their skills. The joint work of practicing scientists and hospital doctors contributes to the growth of their skills, and innovation in a variety of areas of medicine allows us to achieve unique results in the treatment of patients.

Based on the results of the work of City Clinical Hospital No. 40, 9 doctoral and 29 candidate dissertations were defended. More than 70 copyright certificates and patents have been received for methods of operations, diagnostics and utility models developed by hospital specialists.

The work of the team of doctors was highly appreciated by the award of the Nizhny Novgorod City Prize in the field of medicine in 2012. The work is devoted to one of the most pressing and complex problems of surgery - the treatment of phlebothrombosis of the lower extremities, over the last five-year period of observation and provision of emergency and planned surgical care to patients with phlebothrombosis in the conditions of the city phlebological center on the basis of the State Budgetary Institution No. 40.

The maternity hospital, which is part of the hospital, has 205 beds and is the largest in the Nizhny Novgorod region. Currently it serves as a regional perinatal center, accepting 4500-5000 births annually. All work carried out by obstetricians and gynecologists is aimed at reducing the number of cases of miscarriage and post-term pregnancy, achieving natural delivery and improving the survival rates of newborns.

Since January 2010, a Health Center has been successfully operating within the walls of the adult clinic, recognized as one of the best at the regional level. His work experience was presented by the head of the Center at the All-Russian Forum - “A Healthy Nation is the Basis of Russia’s Prosperity.”

In 2012, in order to increase the accessibility of medical care in remote areas, a general practitioner’s office was organized and successfully operates within the structure of the adult clinic.

More than a thousand employees of the institution, including doctors, nurses, laboratory assistants, orderlies, technical staff, with their hard work, care and warmth, create an atmosphere of humanism and mercy, which is characteristic of the staff of the State Budgetary Institution No. 40.

During the internship, the entire process of receiving and registering biomaterial was mastered.

1. Reception of biomaterial and forms.

2. Study of biomaterial

3. Recording the results obtained on a form

4. Recording the results in test logs

5. Disposal of biomaterial

Conclusion

The main form of control over the organization of reception, labeling and registration of biomaterial is periodic external and internal inspections. But this form of control cannot be considered effective. The problem of controlling this stage of laboratory research remains today one of the most serious problems of modern laboratory medicine.

The most effective and efficient step seems to be the creation of standard conditions for the collection, transportation and storage of biological samples from the patient (use of vacuum or other systems). The introduction of such systems into practice does not affect all stages of laboratory research and, in general, takes the organization of laboratory work to a different level. Their use not only facilitates the organization and significantly promotes the standardization of the preanalytical stage, but also creates the conditions necessary for the implementation of modern laboratory systems. If analyzers are available, they can be used as primary tubes, which greatly simplifies the analytical stage of laboratory research.

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Active

MU 4.2.2039-05

METHODOLOGICAL INSTRUCTIONS

4.2. CONTROL METHODS. BIOLOGICAL AND MICROBIOLOGICAL FACTORS

Technology for collecting and transporting biomaterials to microbiological
laboratories

Date of introduction 2006-07-01

2. RECOMMENDED FOR APPROVAL by the Commission on State Sanitary and Hygienic Standards under the Federal Service for Supervision of Consumer Rights Protection and Human Welfare on October 6, 2005 (Protocol No. 3).

3. APPROVED AND ENTERED INTO EFFECT by the Head of the Federal Service for Supervision of Consumer Rights Protection and Human Welfare, Chief State Sanitary Doctor of the Russian Federation G. G. Onishchenko on December 23, 2005.

5. INTRODUCED FOR THE FIRST TIME.

1 area of ​​use

1 area of ​​use

1.1. The guidelines set out the rules for collecting and transporting biological materials to microbiological laboratories in order to improve the quality of laboratory research results and organize anti-epidemic and preventive measures, as well as the prevention of nosocomial infections among medical personnel and patients.

1.2. The guidelines are intended for use by authorities and organizations of the Federal Service for Surveillance on Consumer Rights Protection and Human Welfare, and can also be used by health authorities and organizations.

2. General provisions

2.1. Obtaining reliable data on identifying sources of infection is necessary for the timely and effective organization of anti-epidemic and preventive measures, assessing the level of infection in the population during epidemiological surveillance.

2.2. The developed technique for collecting and transporting biological materials to microbiological laboratories will reduce the level of preanalytical errors and improve the quality of laboratories in objectifying results.

2.4. The guidelines can be used when conducting epidemiological surveillance of antimicrobial resistance of infectious agents isolated and identified in the laboratory, optimizing the use of antimicrobial drugs and measures in the field of prevention, control and containment of resistance at the local, regional and national levels.

3. General requirements for collecting samples of biological material for microbiological research

3.1. To protect medical personnel and patients from infection when collecting samples of biomaterials and delivering them to the laboratory, it is necessary:

do not contaminate the outer surface of the dishes when collecting and delivering samples;

do not contaminate accompanying documents (directions);

minimize direct contact of the biomaterial sample with the hands of the medical worker collecting and delivering it to the laboratory;

use sterile disposable or approved for use for these purposes in the prescribed manner containers (containers) for collecting, storing and delivering samples;

transport samples in carriers or packages with separate nests;

observe aseptic conditions to prevent infection of the patient during the implementation of invasive measures;

collect samples in sterile disposable or glass containers (not contaminated with biomaterial, not damaged by cracks, chipped edges and other defects)

3.2. Biomaterial samples must be collected as follows:

before the start of antibacterial therapy, if this is not possible - immediately before the repeated administration (taking) of drugs;

in the quantity (weight, volume) necessary to perform the analysis, because an insufficient amount of biomaterial for research leads to false results;

with minimal contamination of the material with normal microflora, because its presence leads to erroneous interpretation of the results obtained, for example, from the study of sputum, samples from the nose, pharynx (throat), genitals, etc.

3.3. When collecting a sample, make sure that in the laboratory, when opening the container with the biomaterial, no aerosol is formed: samples of blood and other body fluids are carefully transferred from a syringe without the formation of foam into a dry and/or container filled with a medium (anticoagulant).

3.4. In the direction for the study indicate: last name, first name, patronymic of the patient; year of birth; the department in which he is located; medical history number (outpatient card); diagnosis; material sent for research and research objectives; date and time of material collection (hours); antibacterial (immune) drugs, if the sample is taken against the background of antibiotic and/or immunotherapy; last name, first name, patronymic of the attending physician (consultant) sending the sample for testing. When sending biomaterials obtained during an autopsy, the department in which the patient died is also indicated.

3.5. Before collecting a sample, especially when using invasive methods, the likelihood of risk and benefit to the patient is taken into account, as well as the significance of this particular type of biomaterial for the purpose of objectifying the clinical diagnosis and assessing ongoing or planned therapeutic measures.

4. General requirements for the delivery of biomaterial samples to the microbiological laboratory

4.1. All collected samples are sent to the microbiological laboratory immediately after receipt, with the exception of cases of using containers with transport media approved for use for these purposes in the Russian Federation in the prescribed manner.

This is necessary for:

maintaining the viability of pathogens and the possibility of isolating microorganisms that require special cultivation conditions (Haemophylus, etc.);

preventing excessive growth of fast-growing and active microorganisms;

maintaining the ratio of the initial concentrations of isolates in the presence of microbial associations in the sample;

reducing the time of sample contact with some antiseptics used locally, which may have antibacterial activity;

objectification of the clinical diagnosis of an infectious-inflammatory disease and assessment of the results of therapy.

4.2. It is allowed to use alternative methods to increase the time it takes to deliver biomaterial to the laboratory.

Samples are stored in a refrigerator at a temperature of 2-8 °C, except for the cases listed below.

4.2.1. When the sample is stored in a specialized transport container (transport system), approved for use in the prescribed manner, which is a sterile disposable tube with an agar or liquid transport medium and a swab probe mounted in a stopper and sterilely packaged with the tube. In such containers, samples are stored at room temperature (18-20 °C). Transport media, special dense ones with and without activated carbon, make it possible to ensure the preservation of the viability of microorganisms that require special cultivation conditions for 48-72 hours.

For tests for anaerobes and fecal flora, use special containers with a transport medium, test tubes with media for isolating Campylobacter and Helicobacter, approved for use in the prescribed manner. Such environments create an anabiotic atmosphere for microorganisms, which helps to reduce their metabolism, inhibit growth, and prevent them from drying out and accumulating waste products.

Each sample collected in a liquid medium is thoroughly mixed with the medium.

4.2.2. When blood is cultured in broth, then after obtaining the sample is stored in a thermostat at a temperature of 35-37 ° C.

If samples are collected in special containers for subsequent research with a two-phase environment, they should be stored at room temperature (18-20 °C).

4.2.3. When there is a possible presence of temperature-sensitive microorganisms (Neisseria sp.), the samples are left at room temperature (18-20 °C).

4.3. To transport samples tested for the presence of aerobes and facultative anaerobes, use:

disposable sterile dry tubes with a built-in swab probe (tubes) or containers with a transport medium, approved for use for these purposes in the Russian Federation in the prescribed manner; It is allowed to use sterile glass tubes, sealed with a gas-permeable stopper with a built-in swab probe, prepared in the laboratory;

disposable sterile containers with a screw cap (glass with a gas-permeable stopper are allowed) - for collecting samples of urine, sputum, feces, broncho-alveolar lavage, biopsy (pieces of tissue) material;

sterile disposable with a screw cap or glass tubes - for collecting sterile liquids, broncho-alveolar lavage separated from drainages or scrapings;

sterile Petri dishes - for collecting hair samples or for transporting scrapings with markings on the bottom of the dish;

special sterile nasopharyngeal and urogenital probes-tampons with an aluminum axis (axis diameter - 0.9 mm) and a small cotton or viscose swab at the tip (tampon diameter - 2.5 mm), mounted in a stopper sealing a sterile disposable glass tube - for samples from the nasopharynx for B. pertusis and from the urethra in men.

4.4. To transport samples tested for the presence of anaerobes, containers with special transport media and test tubes with thioglycollate medium are used; test tubes with media for the isolation of Campylobacter and Helicobacter, approved for use for these purposes in the Russian Federation in accordance with the established procedure. The sample collected in a liquid medium is thoroughly mixed with it. The following methods are recommended for obtaining samples:

drainage discharge used for active aspiration of cavities is sucked out with a sterile syringe with a tight piston in a volume of 2-4 ml; a sterile needle covered with a sterile cotton swab is placed on the filled syringe and excess air is removed from the syringe; a cotton swab is dumped into a disinfectant solution; the end of the needle is inserted into a sterile rubber stopper and in this form the syringe with the material is delivered to the laboratory;

the contents of foci of infection and cavities, obtained by puncturing them, are collected in a volume of 2-4 ml using 2-, 5-, 10-ml syringes with a tight piston; excess air is removed from the syringe by covering the needle with a sterile cotton swab, which is then dumped into a disinfectant solution; the needle is disinfected by wiping with a swab moistened with 70% ethyl alcohol; To seal, the end of the needle is inserted into a sterile rubber stopper and in this form the syringe with the material is delivered to the laboratory.

When collecting a large volume of material (3 ml or more), anaerobic bacteria can remain viable for 24 hours at room temperature (18-20 ° C).

If there are only a few drops of discharge, it is transferred from a syringe to a small container or a test tube with transport medium immediately after receipt (containers with transport media are received in the laboratory the day before).

If an anaerobic infection is suspected, pieces of tissue (biopsy material) are collected in sterile disposable containers with a screw cap (allowed in a glass container with a ground-in lid) and delivered to the laboratory immediately.

4.5. To transport samples tested for the presence of viruses, special containers with a liquid medium are used to preserve viruses.

5. Samples of various types of biomaterial and the environment surrounding the patient

Table 1

Glassware used to transport samples to the laboratory

Source and type of clinical material	Products used for sample delivery
	Special transportation containers with a medium, with or without antibiotic neutralizers and reagents that destroy blood cells, approved for use for these purposes in the Russian Federation in the prescribed manner; two-phase medium in bottles
central nervous system
Liquor	Sterile disposable tubes with screw cap; sterile glass tubes with cellulose or cotton-gauze stopper
Material for brain abscesses and biopsy material for inflammatory processes in the central nervous system	A syringe with a needle inserted into a sterile rubber stopper; a test tube with thioglycollate medium, closed with a sterile rubber stopper; transportation containers with a medium for preserving anaerobes
Lower respiratory tract
Biopsy material of the lungs and trachea; sputum, naturally expectorated and induced; scraping from the bronchi	Sterile disposable container with a screw cap (for collecting sputum); laboratory prepared sterile glass container
Tracheal aspirate, broncho-alveolar lavage, bronchial washings	Sterile disposable sputum collection container with screw cap; tightly sealed sterile glass tube
Transtracheal and lung aspirate	Special transportation containers with a medium for anaerobes; a syringe with a needle inserted into a sterile rubber stopper; container with thioglycollate medium; sterile disposable plastic container with a screw cap; tightly sealed tube
Upper respiratory tract
Swabs from the nose, throat, nasopharynx, external ear	A sterile disposable swab probe mounted in a sterile dry tube (tubser), or a transport container with the appropriate medium; transport vessel for viruses; sterile glassware mounted in laboratory
Nasal lavage, nasophageal aspirate	Sterile disposable container with screw cap; transport vessel for viruses
Fluid obtained by tympanocentesis, sinus aspirate obtained by needle aspiration	A syringe with a needle, disinfected after manipulation using a swab moistened with 70% ethyl alcohol, and stuck into a sterile rubber stopper; you can transfer the material from the syringe to a sterile disposable or glass tube or a special vessel with a medium for transporting anaerobes
Tissue obtained during surgery of the nose, pharynx, ear	Sterile disposable container with screw cap; a test tube with thioglycollate or other transport medium, tightly closed with a sterile rubber stopper; tightly closed sterile disposable plastic or glass tube
Scrapings from the conjunctiva of the corner of the eye	Smears on sterile, grease-free glass slides; material that is taken into a special transport container with a medium or inoculated into a nutrient medium
Intraocular fluid	Smears on sterile, grease-free glass slides; material that is taken into a special shipping container with a medium for anaerobes or a syringe with a needle pre-disinfected with 70% ethyl alcohol inserted into a sterile rubber stopper
Discharge taken with a sterile glass rod or a sterile cotton swab from the mucous membrane of the lower transitional fold, from the edge of the eyelids, in case of an ulcer - from the cornea (after anesthesia), in case of angular conjunctivitis - from the corners of the eyelids	A sterile disposable or glass tube with sugar broth, in which a swab probe or a sterile glass rod is installed, used to take a sample; special transport container with virus environment
Secret from the lacrimal sac	A disposable sterile swab probe mounted in a sterile dry tube (tubser) or glass tube
Genitourinary system
Medium amount of freely flowing urine; from the ileal canal used to create an artificial bladder; from a catheter in intensive care patients. Bladder flush. Sample obtained during bilateral urethral catheterization	Sterile disposable urine collection container with screw cap or sterile disposable tube with cap; or a special disposable urine collection tube. When using a sterile glass tube with a cellulose or cotton-gauze stopper, be careful not to soak the stopper with the material (sample volume 10-20 ml)
Sample obtained by suprapubic aspiration	A sterile syringe without a needle, closed with a sterile rubber stopper; a sterile syringe with a needle, previously disinfected with 70% ethyl alcohol and inserted into a rubber stopper
Material from female genital organs
Fluids: amniotic, fallopian tubes, Bartholin	Special transport container with medium for anaerobes; a syringe without a needle, closed with a sterile rubber stopper; a syringe with a needle treated with 70% alcohol and inserted into a sterile rubber stopper (sample volume 1-2 ml)
Samples from the cervical canal, urethra, vagina	A disposable sterile probe-tampon, mounted in a sterile dry test tube (tubeser) or a transport container with a special medium; a swab probe mounted in a cellulose or cotton-gauze stopper of a sterile glass tube. A slide with a prepared smear for the study of bacterial sexually transmitted infections and viruses. Tube with transport medium with activated carbon for preserving gonococci for 48 hours or more
Samples of material from the endometrium	A sterile disposable container with a screw cap or a test tube, or a transport container with a medium for anaerobes; sterile glassware
External genitalia material	A sterile syringe without a needle, closed with a sterile rubber stopper; a slide with a smear covered with a coverslip for determining T. pallidum; a swab probe mounted in a sterile disposable tube (tube) or glass tube; a special transport tube with a medium with activated carbon for preserving gonococci and transport containers for viruses and chlamydia; smears on glass slides to detect other sexually transmitted infections
Material from male genital organs
Urethral swabs	A probe-tampon on an aluminum axis (urethral probe-tampon), mounted in sterile disposable (tubser) or glass tubes; a special transport tube with a medium with activated carbon for preserving gonococci and transport vessels for viruses and chlamydia; smears on glass slides to detect other sexually transmitted infections
Ejaculate, sperm	Sterile disposable container with screw cap; sterile tube or tube, disposable or glass
Material of the epididymis in epididymitis	A special shipping container with a medium for anaerobes or a container with a thioglycollate medium; a sterile disposable container with a screw cap or a sterile glass tube with a cellulose or cotton-gauze stopper
Penis lesion	A syringe without a needle, closed with a sterile rubber stopper; a slide with a smear of T. pallidum covered with a coverslip; a swab probe mounted in a sterile disposable tube (tube) or glass tube; shipping containers for viruses and chlamydia; swabs to detect other sexually transmitted infections
Material for suspected gonorrhea
Smears from the anus, cervical canal, urethra, vagina	A probe-tampon mounted in a sterile disposable (tubeser) or glass tube; special transport tube with activated carbon medium to preserve gonococci for 48 hours or more
Gastrointestinal tract
Oral cavity	A probe-tampon mounted in a sterile disposable (tubeser) or glass tube; sterile disposable or glass container for collecting oral fluid
Gastric lavage or lavage fluid; duodenal aspirate; sample obtained during sigmoidoscopy; rectal biopsy material	Sterile disposable container with screw cap; a special sterile container for collecting sputum; sterile glass container; aspirate - in a sterile syringe with a needle, previously disinfected and inserted into a sterile rubber stopper
Rectal swab	A disposable sterile probe-tampon, mounted in a sterile dry test tube (tubeser); or a sterile tube with a special transport medium; probe-tampon made of stainless material, mounted in a cellulose or cotton-gauze stopper of a sterile glass tube
Feces for the presence of intestinal infections ("disgroup"), Helicobacter, Campylobacter	Sterile tube with a swab in saline solution with glycerin; transport container with a medium for anaerobes in fecal samples, with special media with and without activated carbon for the isolation of Campylobacter sp. and Helicobacter sp.; probe-tampon in a dry, sterile disposable tube (tubeser); sterile glass tube mounted with a swab probe on a metal wire made of titanium, steel, aluminum. It is not allowed to use a tampon with a wooden axis
Feces for the presence of dysbacteriosis according to the generic and species composition of microbes (“for flora”)	Special transport container with media for anaerobes in fecal samples, with and without activated carbon media for the isolation of Campylobacter sp. and Helicobacter sp., approved for use for these purposes in the Russian Federation in accordance with the established procedure; probe-tampon in a test tube with saline solution with glycerin in a sterile disposable or glass tube; a special disposable sterile container with a screw cap; laboratory-mounted glass container
Feces for the presence of dysbacteriosis with quantitative accounting of isolated and identified microbes (“for dysbacteriosis”)	Special sterile disposable containers with a screw cap and a spatula for collecting material and taking a sample for culture, having a standard weight; special sterile glass container mounted in the laboratory
Skin and subcutaneous tissues
Ulcers, nodules (nodular thickenings), shallow, superficial wounds (purulent; burns); deep wounds or abscesses, bones	Special sterile disposable containers with a screw cap; sterile tubes with stoppers, disposable or glass, 5 ml; containers with special media for anaerobes
Exudate of subcutaneous and soft tissues; soft tissue aspirate	A sterile syringe without a needle, closed with a sterile rubber stopper; a syringe with a needle, previously disinfected with 70% ethyl alcohol and stuck into a rubber stopper
Sterile body fluids, excluding blood, cerebrospinal fluid, urine (see above)
Liquids
Pleural, peritoneal, ascitic, articular, synovial	Sterile disposable container with screw cap; a closed syringe without a needle or with a needle, previously disinfected with 70% ethyl alcohol and inserted into a sterile rubber stopper
Biomaterial for PCR diagnostics
Blood; other body fluids	Sterile, disposable, stoppered 1.5 ml tubes with anticoagulant. The tube is inverted 3-5 times to mix the sample with the anticoagulant. Delivery to the laboratory - in a rack made of materials that can be sterilized in an autoclave
Biopsy material; scrapings; sputum and other types of biomaterial	Sterile dry disposable Eppendorf tubes and other similar ones. Delivery to the laboratory - in a special rack made of materials that can be sterilized in an autoclave
Samples of the patient's environment
Air	Sterile Petri dishes, disposable (=90 mm) or glass (=100 mm) with dense nutrient media. Delivery to the laboratory in special carrying containers
Washings from environmental objects surrounding the patient
Tests to determine sterility
Blood and blood components from the collection department	Containers with prepared material are delivered to the laboratory in special carrying containers
Flushes from medical instruments, hoses of equipment used in intensive care and anesthesiology departments, as well as in operating rooms; hands of medical personnel; linen	Sterile swab probes mounted in transparent tubes containing a clear, colorless liquid medium. Samples are delivered to the laboratory in racks made of materials that can be sterilized in an autoclave.
Surgical material: tampons, napkins, turundas, cotton swabs on a wooden axis	Sterile transparent test tubes with stoppers containing a clear colorless liquid medium with 3-5 sterile glass beads. Samples are delivered to the laboratory in racks made of materials that can be sterilized in an autoclave.
Suture material: catgut stored in the operating room in an alcohol solution of iodine	Sterile test tube (bottle with stopper) with neutralizer solution (sodium hyposulfite)
Silk, nylon, monomer synthetic threads stored in an alcohol solution in the operating room	A sterile test tube with a stopper containing sterile distilled water as a wash liquid. Suture material samples are delivered to the laboratory in racks that are subject to sterilization in an autoclave.
Washings from the patient's surgical field	Before processing the field: into transparent tubes filled with a clear liquid nutrient medium. After processing the field: a sterile test tube (bottle with a stopper) with a neutralizer solution (sodium hyposulfite). The material is delivered to the laboratory in racks that are subject to sterilization in an autoclave.
Tests to determine the immunological factors of the anti-infective defense system (system of anti-infectious resistance of the body - SAIR)
Blood and other body fluids to determine humoral protective factors	Disposable syringes-test tubes (vacutainers); sterile disposable tubes with screw cap; sterile glass tubes with sterile rubber, cellulose or cotton-gauze stoppers. Samples are delivered to the laboratory in a special rack made of materials that can be sterilized in an autoclave. When using glass test tubes with cotton-gauze stoppers, do not soak them during transportation.
Blood and other body fluids to determine cellular protective factors	Sterile disposable tubes with screw cap; sterile glass tubes tightly closed with a sterile rubber stopper. Disposable tubes with anticoagulant, subject to centrifugation. Delivery to the laboratory is carried out in a special rack made of materials subject to sterilization in an autoclave.
Smears to determine bacterial and viral antigens (for example, if sexually transmitted infections are suspected)	Sterile, defatted glass slide; for T. pallidum, the slide must be covered with a coverslip. Delivery to the laboratory is carried out in a disposable container or sterile disposable or glass Petri dishes in special containers

Note. Specimens of pus, fluid, and tissue should be transported to the laboratory in special, sterile, disposable containers with screw caps. Probes-sampons - dry in sterile disposable tubes (tubes) or in containers with a transport medium, if the material cannot be delivered to the laboratory immediately after taking the sample. For these purposes, you can use transport media, incl. in disposable tubes, packaged with a sterile swab probe, approved for use in the Russian Federation in accordance with the established procedure. Probe-swabs are produced (prepared) using a number of materials: for the swab itself (cotton-gauze, cotton, viscose, dacron), for the axis of the probe-swab (wood, plastic, stainless metal - titanium, steel, aluminum) and dense agar media : with and without coal for anaerobes.


Many clinically significant anaerobes, for example, Clostridium perfringens, Bacteroides fragilis, are quite tolerant to the presence of oxygen and are well preserved in large quantities of pus, samples of fluids and body tissues, as well as on a swab probe in special transport or thioglycollate media.

More sensitive to oxygen (for example, fusobacteria) are also well preserved in a sample of biological material, and if the material is inoculated within 2-3 hours from the moment of its collection, then no special attention is required to the delivery method. Anaerobes that require special cultivation conditions do not remain viable on a swab probe in a transport container with a medium not intended for the delivery of anaerobes, with the exception of containers with special transport media or thioglycollate.

At the same time, these microorganisms grow 5-7 days longer than the time during which most practical laboratories incubate dishes seeded with material for the isolation and identification of aerobes and facultative anaerobes. For this reason, in ordinary practical laboratories, it seems impractical to routinely test for the presence of anaerobes using special transport anaerobic containers with media, and one can limit ourselves to tubes with thioglycollate medium.

To collect and transport samples, use media, containers, tools and materials approved for use in the Russian Federation in accordance with the established procedure.

6. Rules and techniques for obtaining samples of various types of biological material for microbiological research

It is recommended that any sample that requires the use of invasive methods be obtained from a patient by a physician (with the exception of a blood sample that can be collected by a procedural nurse).

If it is necessary to collect material using a swab probe on an open surgical surface or in case of other invasive intervention, it is permissible to use ready-made swab probes with an international safety level of at least Class IIA, approved for use for these purposes in the Russian Federation in the prescribed manner.

6.1. Blood samples

Samples to determine the presence of biological agents in the blood (bacteremia, viremia, etc.) are obtained by venipuncture of peripheral veins (usually the veins of the elbow), arteries, or from the heel of newborns.

Collection of a sample from an indwelling intravenous or intra-arterial catheter is allowed only in cases of suspected catheter-associated infection or the possibility of obtaining it by venipuncture.

6.1.1. In acute sepsis, meningitis, osteomyelitis, arthritis, acute untreated bacterial pneumonia and pyelonephritis, 2 samples are collected from two vessels or two sections of a blood vessel before starting antibacterial therapy.

6.1.2. If you suspect the presence of endocarditis and sluggish sepsis with a small (10-30 CFU/ml) concentration of the pathogen in the circulation:

in the presence of an acute process, 2 samples are collected from two sections of blood vessels (different vessels) during the first 1-2 hours of rising body temperature (not at the peak of temperature!) and before the start of therapy;

in case of subacute or sluggish course, 3 samples are collected on the first day with an interval of 15 minutes or more. If all samples are negative, 3 more are collected on the second day after sowing;

in patients with endocarditis receiving antibiotics, 2 separate samples are collected on each of three days with positive clinical dynamics of therapy;

to confirm the clinical diagnosis of “infective endocarditis”, if it is impossible to obtain the required number of blood samples from the patient for microbiological analysis or if the culture results are negative, blood samples are sent to study the immunological factors of the body’s defense system (anti-infective resistance system - SAIR); delivery to the laboratory is carried out in sterile glass or disposable test tubes with a cap (stopper), received the day before in the laboratory;

if the patient suffers from congenital heart disease (CHD), it should be remembered that CHD is the prodrome of infective endocarditis, and the examination algorithm for such a patient includes the determination of immunological factors of CAIR.

6.1.3. In patients whose therapy includes antibiotics, 6 samples are collected within 48 hours; samples must be collected immediately before administering (taking) the next dose of the drug.

6.1.4. If a patient has a fever of unknown origin, initially 2 samples are collected from different blood vessels (two sections of the vessel), then after 24-36 hours, 2 more samples are collected against the background of an increase in body temperature (not at the peak of the temperature!).

6.1.5. Technique for obtaining a blood sample. Blood samples for culture are collected by 2 people at the patient's bedside or in the treatment room.

To obtain a sample, you must do the following:

disinfect the area of ​​skin above the vessel selected for puncture: treat the skin with a swab moistened with 70% ethyl alcohol, then with another swab moistened with a 1-2% iodine solution or other disinfectant approved for use for these purposes in the prescribed manner, in a circular motion , starting from the center, for 30 s;

wait until the treated area dries. It is not allowed to palpate the vessel after treating the skin before inserting the needle;

when working with vials with dual media: use a sterile syringe to collect 10 ml of blood from adults, 5 ml from children; open the bottle over the flame of an alcohol lamp; add blood into the vial from a syringe, after removing the needle; burn the neck and stopper of the bottle in the flame of an alcohol lamp, close the bottle with the stopper; Carefully, so as not to soak the bottle cap, mix its contents in a circular motion.

When using ready-made bottles with media and reagents that neutralize antibiotics and destroy blood cells, or without them, approved for use for these purposes in the Russian Federation in accordance with the established procedure, 10-30 ml of blood is obtained from adults, 0.5-ml from children. 3.0 ml.

Wherein:

in parallel with the disinfection of the skin area for puncturing, the bottle caps are treated with 70% ethyl alcohol (iodine solution is not allowed to be used to treat the caps when working with bottles, for example, Bactec, Vital and other similar ones, approved for use for these purposes in the Russian Federation in accordance with the established order);

blood received from adults in equal volumes, after puncturing the cap of the container, is introduced into the “aerobic” and “anaerobic” containers; blood received from children - into a special “baby” bottle, piercing the cap of the container.

6.1.6. After venipuncture and blood culture in a container with a medium to prevent possible irritation (burn), the remaining iodine is removed from the patient’s skin using a swab moistened with 70% ethyl alcohol.

6.2. Tests for infectious and inflammatory processes of the central nervous system (CNS)

6.2.1. Liquor. 4.0-5.5 ml of cerebrospinal fluid obtained during lumbar puncture from the subarachnoid space between the vertebrae L3-L4, L4-L5 or L5-S1, as well as during puncture of the lateral ventricles of the brain, is sent to the microbiological laboratory for research.

Sample collection is carried out by slowly filling three tubes with three portions of material for research in laboratories. Use sterile tubes with tight-fitting caps (disposable with a stopper or glass with a sterile rubber stopper).

Of the three tubes with material obtained during lumbar puncture, the tube with the most turbid content is always sent, as a rule, this is the second tube in the sample collection process.

When obtaining the material by puncture of the lateral ventricles of the brain, freshly taken cerebrospinal fluid from the syringe, having previously removed the needle, is introduced into a sterile test tube over the flame of an alcohol lamp, the neck of the tube and the stopper are burned in the flame of the alcohol lamp (when working with a glass test tube sealed with cotton-gauze or rubber stoppers), and closed test tube with a stopper.

In all cases suspected of meningitis, in addition to the cerebrospinal fluid, material is collected from the suspected foci of infection: swabs from the nasopharynx, middle ear, blood samples, and together with the cerebrospinal fluid they are sent to the laboratory. A good way to preserve meningococcus for a long time (up to 48 hours) is to take the material with a smear with a swab probe and place it in a test tube with a transport medium with or without activated carbon.

Liquor for microbiological research is immediately sent to the laboratory on a heating pad to maintain a temperature of 35-37 °C. If this is not possible, the liquor is collected in a container with a transport medium and left in the refrigerator (at a temperature of 4-8 ° C) until the morning, and then delivered to the laboratory.

If it is necessary to conduct virological studies, cerebrospinal fluid samples are placed in a refrigerator at a temperature of 2-8 ° C or frozen, or stored at room temperature using containers with a special liquid medium.

6.2.2. Material from brain abscesses. Considering that in 90% of cases anaerobes grow in the sample, the material is aspirated from the outbreak and sent to the laboratory in a container with an anaerobic environment, approved for use for these purposes in the Russian Federation in accordance with the established procedure, or in the syringe with which the sample was collected, having previously removed the needle and closing the syringe with a sterile rubber stopper.

The sample is delivered immediately upon receipt.

6.2.3. Biopsy material. Samples are obtained during surgery and placed in a container with an anaerobic environment or in a sterile tube with thioglycollate medium, closed with a sterile rubber stopper.

The material is sent to the laboratory immediately.

6.3. Tests for infectious and inflammatory processes of the eyes

Most samples obtained from the eye are collected by an ophthalmologist. These samples should be cultured on culture media at the patient's bedside or in the treatment room, or in the doctor's office during an appointment, and the inoculated material should be transferred to the laboratory for cultivation, isolation, identification and determination of antibiotic sensitivity of pathogens. Samples taken using invasive and other aggressive methods are collected in parallel with a conjunctival smear, which in such cases serves as a control.

The day before, 6-8 hours (night), all medications and procedures are canceled.

If there are specific clinical manifestations of an infectious-inflammatory process or the doctor has suspicions, before anesthesia it is necessary to prepare smears to determine chlamydia and viruses, and send the drugs to the laboratory. To preserve chlamydia and viruses, the material is collected in containers with transport media.

The discharge is collected with a sterile glass rod or a sterile swab probe: rubbed in two or three movements along the mucous membrane of the lower transitional fold, from the edge of the eyelids; for an ulcer - from the cornea (after anesthesia), for "angular conjunctivitis" - from the corners of the eyelids.

The secretion from the lacrimal sac is collected with a sterile swab probe after gentle massage.

The material taken with a stick and/or swab probe is placed in a container with a transport medium or in a sterile glass tube with a liquid medium and delivered to the laboratory, being careful not to soak the cellulose or cotton-gauze stopper of the tube.

When using swab probes, it is preferable to use disposable containers with transport medium or sterile glass tubes with cotton-gauze stoppers filled with sterile medium.

The most accurate information about the causative agent of the inflammatory process can be obtained from microbiological analysis of scrapings.

6.3.1. Conjunctivitis and blepharoconjunctivitis. The day before, 6-8 hours (night), all medications and procedures are canceled for patients in the hospital.

Conjunctival samples are collected using a sterile, pre-moistened viscose or calcium alginate swab probe located in a disposable sterile tube or glass tube. Samples from each eye are collected with separate swabs using two or three circular movements along the mucous membrane.

Tubes with swabs from each eye are labeled “right” and “left”, respectively, and immediately sent to the laboratory.

Before obtaining a scraping, 1-2 drops of an anesthetic are administered, for example, proparacaine hydrochloride or another that is available and approved for use for these purposes in the prescribed manner.

Using two or three short sharp movements in one direction, scrapings from the conjunctiva are collected using a special sterile spatula. During the manipulation, the eye must be open.

Be careful not to touch your eyelashes when collecting the sample.

Prepare at least 2 smears from each eye by applying the material to a clean, grease-free glass slide in a circular motion over a 1 cm diameter area.

Fix the smears for 5 minutes in 95% methyl alcohol in a special, tightly closed container.

Immediately test tubes with swab probes and glasses with smears are transferred to the laboratory.

6.3.2. Bacterial keratitis. The day before, 6-8 hours (night), all medications and procedures are canceled for patients in the hospital.

Receive 2 samples from the conjunctiva, as described in paragraph 6.3.1, because the results of their culture may be useful in determining the source of corneal contamination (one sample is used if a fungal infection is suspected).

If a viral infection is suspected, the conjunctival exudate and the material obtained by scraping are placed in a special transport medium for viruses.

Anesthesia is carried out as indicated in paragraph 6.3.1.

Collect 3-5 scrapings using the method described in paragraph 6.3.1.

Place the collected material in a small (5 ml) sterile disposable screw cap tube or sterile glass tube with a rubber stopper filled with medium.

Prepare 2-3 smears on a glass slide, and after drying, fix them as described in paragraph 6.3.1.

Immediately transfer all material to the laboratory.

6.3.3. Bacterial endophthalmitis. A sample of vitreous fluid (1-2 ml) is collected by fine needle aspiration and a vitrectomy sample is obtained.

Next do the following:

From the syringe, after removing the needle, the contents are placed in a disposable sterile container with a screw cap or a test tube tightly closed with a rubber stopper;

the material sample can be left in the syringe by removing the air and removing the needle, closing the syringe with a sterile rubber stopper;

collect a sample from the conjunctiva as described in paragraph 6.3.1;

all material immediately sent to the laboratory.

6.3.4. Preseptal cellulitis. Treat the skin with 70% ethyl alcohol and 1-2% iodine solution or iodoform or other available disinfectant approved for use in the prescribed manner.

If there is no open wound, the upper or lower eyelid is pierced to collect material.

If there is an open wound, collect a sample of purulent material with a syringe and needle.

Prepare smears on a glass slide as described in paragraph 6.3.1.

Transfer the collected material into a transport container with media for anaerobes or leave it in a syringe, which, after removing the air from it and removing the needle, is closed with a stopper.

Collected material and smears immediately sent to the laboratory.

6.3.5. Orbital cellulitis. Collect aspirate from the zone according to the method described earlier in paragraph 6.3.2.

Collect a sample from the conjunctiva and prepare smears on a glass slide, as in paragraph 6.3.1.

For delivery to the laboratory, use the materials described in clause 6.3.3.

Blood samples are collected from the patient according to the method described in paragraph 6.1. Blood samples and all collected material are sent to the laboratory.

6.3.6. Inflammation of the lacrimal gland (dacryoadenitis).

Collect purulent contents using a probe-tampon, as in paragraph 6.3.1.

Do not use the needle aspiration method, so as not to disrupt the integrity of the lacrimal gland.

The swab probe and slides with smears are sent to the laboratory.

6.3.7. Inflammation of the lacrimal sac (dacryocystitis). Obtain a sample from the conjunctiva, as in paragraph 6.3.1.

Massage and press on the lacrimal sac to obtain a sample of exudate for culture and preparation of smears or, using another method, collect exudate with a syringe and needle.

Place the collected exudate sample in transport containers as described earlier and transport it to the laboratory.

6.3.8. Canaliculitis. Press on the inside of the eyelid to extract pus.

Next, work is carried out according to the methods outlined earlier in this section.

If there is a specialized department (clinic) in a treatment-and-prophylactic organization, all cultures are performed at the patient’s bedside using nutrient media obtained in the laboratory. The inoculated material along with the prepared smears is transferred to the laboratory.

6.4. Tests for infectious and inflammatory processes of the ear

6.4.1. If the outer ear is affected, the skin is treated with 70% alcohol, followed by rinsing with sterile saline. The discharge from the lesion is collected on a sterile disposable probe-tampon of a tubeser or test tube with a transport medium, or on a tampon mounted in a sterile glass tube, closed with a stopper, with or without a special medium. The swab is placed in a tube and delivered to the laboratory.

6.4.2. If the middle and inner ear are affected, punctates and other material obtained during surgery are collected. The items are delivered to the laboratory in a closed syringe with the air previously removed. Tissue samples - in a transport container with a medium for anaerobes, approved for use for these purposes in the Russian Federation in accordance with the established procedure, or in a sterile disposable container with a screw cap. It is possible to use a sterile glass tube closed with a sterile rubber stopper.

6.4.3. Tympanocentesis of the tympanic membrane is performed for microbiological diagnosis of middle ear infections only in cases where the patient does not respond to previous therapy or in the torpid course of catarrhal otitis media, even in the visual absence of exudate in the protruding blood (seeds from the nasopharynx are positive in less than 90% of cases).

To obtain a sample, clean the external canal using a swab moistened with 70% ethyl alcohol, followed by treatment with sterile saline.

Using a syringe, collect fluid from the tympanic cavity. The sample is delivered to the laboratory in a sterile disposable container with a screw cap or in a closed syringe with previously removed air.

If the eardrum is damaged, the material is collected with a swab probe using a mirror. The swab is placed in a sterile disposable tube (tubser) or container with a transport medium (a sterile glass tube with or without medium is allowed), and in this form is delivered to the laboratory.

6.5. Tests for infectious and inflammatory processes of the respiratory tract

6.5.1. Upper respiratory tract. If a patient is suspected of having diphtheria, whooping cough, chlamydia, mycoplasmosis, legionellosis, or gonorrhea, laboratory workers are informed before the sample is delivered so that they are prepared to analyze this type of material.

6.5.1.1. A sample from the mucous membranes of the anterior sections of the nasal cavity is collected with one sterile swab probe mounted in a sterile disposable tube (tubeser) or specially mounted in a sterile glass tube:

remove the tampon from the test tube, insert it into the right nostril and, using rotational movements, collect material from the wings of the nose and the upper corner of the nasal opening;

repeat the manipulation for the left nostril;

The swab is placed in a test tube and delivered to the laboratory.

Samples obtained in this way are useful for determining the carriage of Staphylococcus aureus in medical workers, the presence of pathogens of nosocomial infections, as well as for characterizing dysbiotic disorders of the mucous membranes of the upper respiratory tract during a comprehensive clinical immunomicrobiological examination of patients carried out in scientific and practical laboratories.

If there are foci of inflammation or ulceration in the nasal cavity, material from the lesion(s) is collected with a separate swab.

6.5.1.2. Nasopharyngeal aspirates are collected to determine the carriage of streptococcus pyogenes, meningococcus, diphtheria and pertussis pathogens, as well as for epidemiological studies of antimicrobial resistance of S. pneumoniae and H. influenzae:

suck out material from the nasopharynx;

transfer the material into a sterile disposable container with a screw cap or a special sterile tube with a gas-permeable cellulose or cotton-gauze stopper, being careful not to soak it with a sample of the clinical material.

6.5.1.3. Nasopharyngeal swab. The material is collected to determine the carriage of meningococcus and diagnose whooping cough.

Using gentle rotational movements along the lower nasal passage, a cotton, viscose or calcium alginate probe-tampon is inserted into the nasopharynx alternately into both nostrils. At the same time, the wings of the nose are pressed against the tampon and the nasal septum for closer contact with the mucous membrane.

The swab is placed in a sterile tube and delivered to the laboratory.

6.5.1.4. Nasal lavage is used to determine the presence of not only a bacterial, but also predominantly a viral infection.

The patient is warned not to swallow during the procedure.

Place the patient's head in an up-and-down position at an angle of approximately 70°, inject 5 ml of sterile saline into each nostril, the patient should remain in this position for 3-5 seconds.

To collect material, lower the patient's head forward to allow fluid to flow from the nostrils into a sterile disposable container, or aspirate the fluid by inserting a rubber drain into each nostril.

Place an equal volume of the wash into a container containing viral transport medium or deliver this portion in a sterile disposable container.

6.5.1.5. Sinus punctate. Using a syringe aspiration technique, a specially trained, experienced physician or otolaryngologist obtains material from the maxillary frontal or other sinuses. The contents of the syringe are placed in a container with transport medium for anaerobes or in a sterile tube with thioglycollate medium. You can leave the material in the syringe and, closing it with a sterile rubber stopper, deliver it to the laboratory.

6.5.1.6. Obtaining a sample from the mucous membrane of the pharynx (pharynx).

It is not allowed to collect material from the pharynx (pharynx) with an inflamed epiglottis, since the procedure can lead to serious respiratory obstruction.

When taking a sample from the mucous membrane of the pharynx (pharynx), do not touch the mucous membranes of the cheeks, tongue, gums, lips with a swab, and do not collect saliva, since this material characterizes the mucous membranes of the oral cavity, that is, the upper part of the gastrointestinal tract.

A swab from the throat (pharynx) is collected on an empty stomach or 3-4 hours after eating. Before taking a sample, the patient should rinse his mouth with warm boiled water.

To obtain a sample, use a sterile spatula or swab: remove a viscose swab from a sterile disposable test tube (tube) or use a swab prepared in the laboratory, mounted in a sterile glass test tube. The use of a swab probe with a viscose head is preferable, because viscose adsorbs less liquid and more cellular material.

With one hand, press the patient's tongue with a sterile spatula.

With the other hand, they collect the material, alternately treating the right tonsil, the right palatine arch, the left tonsil, the left palatine arch, and the uvula with a tampon; at the level of the uvula, they touch the posterior wall of the pharynx with the tampon.

Samples obtained in this way are useful for determining the presence of a pathogen of nosocomial infection, as well as for characterizing dysbiotic disorders of the mucous membranes of the upper respiratory tract during a comprehensive clinical and immunomicrobiological examination of patients carried out in scientific and practical laboratories.

If there are foci of inflammation or ulceration on the mucous membrane, be especially careful when collecting the sample and collect additional material from the lesion(s) with a separate swab.

Place the swab in a sterile disposable or glass tube and deliver it to the laboratory.

6.5.1.7. If diphtheria is suspected, laboratory workers are informed, indicating the diagnosis in the direction.

In cases of a respiratory pathological process, material is simultaneously collected from the mucous membranes of the nasopharynx and pharynx according to the methods presented earlier.

If a cutaneous form of diphtheria is suspected, material is collected from the skin, as well as from the mucous membranes of the pharynx (pharynx) and nasopharynx, and all samples are delivered to the laboratory.

6.5.2. Lower respiratory tract. Microbiological diagnosis of inflammatory processes in the lower respiratory tract presents serious difficulties, because During the collection process, the sample may be contaminated with microorganisms that colonize the upper respiratory tract. This ecological niche is quite abundantly contaminated with opportunistic microbes, especially in the presence of dysbacteriosis (qualitative - species composition and/or quantitative - concentration of species present in CFU/ml in a weakened and/or immunocompromised contingent of subjects).

For this reason, samples of material from the lower respiratory tract are collected with particular care to obtain correct information about the etiological agent(s). When using invasive methods to collect a sample (if this does not occur during surgery), the instruments used pass through the upper respiratory tract, and there is a real possibility of seeding deeper loci with microorganisms that inhabit the upper respiratory tract.

6.5.2.1. Sputum.

Freely expectorated sputum- morning collection is preferred.

Before collecting the sample, the patient, if possible, should brush his teeth and rinse his mouth and throat with warm boiled water; if the patient is not able to do this himself, then the toilet of his oral cavity is carried out by medical workers.

The patient is warned not to collect saliva or nasopharyngeal discharge into the container.

A sputum sample obtained as a result of a deep cough is collected in a special sterile disposable container with a screw cap or in a specially prepared sterile glass jar.

The sputum sample is sent to the laboratory.

Induced sputum (recommended mainly if Mycobacterium tuberculosis and Pneumocystis yiroveci are suspected)- morning collection is preferred.

Before collecting the sample, the patient, if possible, should brush his teeth and rinse his mouth and throat with warm boiled water; If the patient is not able to do this himself, then the toilet of his oral cavity is carried out by medical personnel.

Before the procedure, moisten a clean toothbrush with warm boiled water and brush it over the mucous membranes of both cheeks, tongue and gums.

Actively rinse the patient's mouth with warm boiled water.

Using an inhaler, allow the patient to swallow 20-30 ml of 3-10% sterile saline solution.

Collect induced sputum in a special sterile disposable container with a screw cap or in a sterile glass jar prepared accordingly.

The sample is sent to the laboratory.

Simultaneously with the sputum sample, a sample from the pharynx (throat) should be sent to the laboratory, collected after toileting the oral cavity and immediately before collecting sputum, freely separated (expectorated) or induced.

6.5.2.2. Tracheostomy and endotracheal aspirates. The tracheostomy is colonized by microorganisms within 24 hours after the patient's intubation, as a result of which the results of the cultural study have low clinical significance. Given the above, culture results obtained in intubated patients must be constantly compared with clinical data (for example, fever or the appearance of infiltrates on x-ray).

Bronchial lavage or bronchoalveolar lavage samples are collected, if possible, before scraping or biopsy samples are obtained. The rule is dictated by the need to avoid excess blood in the resulting fluid, because blood can change the concentration of cellular and non-cellular components of the sample and affect the result of microbiological analysis.

The aspirate sample is collected in a sterile disposable container with a screw cap, or in a sterile glass container mounted appropriately, or in a sterile disposable glass tube with a stopper, or delivered to the laboratory in a closed syringe with the air removed.

6.5.2.3. Samples obtained using a bronchoscope. Bronchoalveolar lavage (sample of choice), bronchial lavage (low sensitivity for diagnosing pneumonia), bronchial scraping (more significant than lavage), transtracheal biopsy samples are obtained by introducing a bronchoscope transnasally or transorally in a non-intubated patient or through an endotracheal tube in an intubated patient.

To obtain a bronchial lavage or bronchoalveolar lavage sample:

sterile non-bacteriostatic (official) physiological solution is injected with a syringe through the biopsy channel of the bronchoscope in separate portions (total volume from 5-20 to 100 ml);

Before introducing the next portion of saline solution, carefully aspirate the inserted part of the syringe into a sterile disposable container with a screw cap or into a sterile disposable or glass tube with a stopper, or leave it in a closed syringe, having previously removed the air from it (usually 50-70% of the injected saline solution is in lavage);

each sucked portion is collected in a separate container;

At the end of the procedure, samples obtained from the same area are combined. Samples from different sites (eg, the right upper lobe of the lung and the right lower lobe) should be combined together only after consultation with the treating physician;

in the direction indicate the total volume of injected saline solution.

To obtain a bronchial scraping sample:

a telescopic double catheter with the distal end treated with polyethylene glycol (or other appropriate reagent) is inserted through the biopsy channel of the bronchoscope to prevent contamination of the sample;

collect the material in a sterile disposable container with a screw cap or in a transport container with an anaerobic medium or in a sterile tube with thioglycollate medium tightly closed with a sterile rubber stopper;

deliver the material to the laboratory.

To obtain a transbronchial biopsy, a sample is collected through the biopsy channel of the bronchoscope and, placing it in a sterile disposable container with a screw cap with a small amount (1-2 ml) of non-bacteriostatic (official) saline solution or in a test tube with thioglycollate medium, tightly closed with a rubber stopper, is transferred to laboratories

6.5.2.4. Lung aspirate samples. To collect the sample, a needle is inserted through the sternum into the lung infiltrate under the control of a computed tomography scanner. The material is aspirated from the site of inflammation. If there is a large infiltrate or several of them, it is necessary to obtain several samples from the corresponding lesions or several samples from one large lesion. The material is transferred to the laboratory in a transport container with anaerobic medium or in a glass tube with thioglycollate medium, or in a screw-on disposable container.

6.5.2.5. Lung biopsy samples. If possible, obtain pieces of tissue 1-3 cm in size. If the lesion is large or there are several of them, several samples are collected. Place the sample in a sterile disposable container with a screw cap or in a transport container with a medium for anaerobes, or in a container (test tube) with a thioglycollate medium closed with a sterile rubber stopper.

6.6. Tests for infectious and inflammatory processes of the genitourinary system

6.6.1. Urine samples.

6.6.1.1. General rules for sample collection:

It is not allowed to collect urine from bed linen or from a urine bag;

to analyze urine during natural urination, use the average morning portion;

before collecting the sample, it is necessary to thoroughly rinse the external genitalia and anal area with warm boiled water; In this case, special attention should be paid to the treatment of the opening of the urethra in men (the vestibule of the vagina in women) to ensure that during the procedure the sample will not be additionally contaminated with microbes;

It is not allowed to use disinfectants for processing, because when introduced into a sample, they can inhibit the growth of microorganisms;

the sample is transferred to the laboratory no later than 2 hours from the moment of collection;

To collect and deliver samples to the laboratory, use sterile disposable containers with a screw cap or test tubes (it is possible to use sterile specially mounted glass tubes with a gas-permeable cellulose or cotton-gauze stopper, but special attention is paid to not soaking the stopper);

delivery to the laboratory of aspirate samples obtained using the technique of suprapubic puncture of the bladder is carried out in a transport container with a special medium for anaerobes or in a closed syringe without a needle;

if a viral infection is suspected, samples are delivered to the laboratory in a sterile disposable container placed on melted ice (dry ice is not allowed!);

any procedure for obtaining a sample using a catheter (especially in women) is carried out with careful adherence to the rules of asepsis to prevent additional contamination of the patient during the insertion of the catheter;

For microbiological research, it is not allowed to use a sample from a daily sample.

6.6.1.2. Collecting a urine sample from women during natural urination.

Use a medium portion, because... the first passage of urine should remove any commensals that may be present in the urethra. The sample collected during the following passages must be free of contamination. The reliability of positive results from such material is 80% when collecting one sample, 90% when collecting two consecutive samples, and 100% when collecting three samples if all samples give the same results.

The medical worker collecting the sample should wash their hands with soap, rinse with water, and dry. If the sample is collected by the patient himself, he is explained in detail how this is done, taking into account the preparation outlined above, and is warned about what may happen if he breaks the rules:

you should thoroughly rinse the opening of the urethra and the area of ​​the vestibule of the vagina, as well as the perineum and the anus with soapy water or liquid soap, rinse with warm boiled water, and dry with a sterile gauze cloth;

the vaginal opening must be closed with a sterile cotton swab;

keep the outer labia at a distance from each other during urination;

drain a small amount of urine into a special container for disposal, while continuing to urinate;

Collect a medium amount of urine (10-20 ml) in a special disposable container with a screw cap or a sterile glass container specially installed in the laboratory.

6.6.1.3. Collecting a urine sample from men during natural urination.

In men, one portion of urine is required for microbiological examination, because their likelihood of contamination is significantly less than that of women.

The person collecting the sample should wash their hands with soap, rinse with water, and dry. If samples are collected by the patient themselves, they should be explained in detail how this is done, taking into account the preparation outlined below, and warned of what may happen if the rules are not followed:
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Carrying out research using the PCR method is allowed on the basis of existing laboratories, provided that the laboratory has independent work areas or work areas allocated as part of other functional premises that correspond to the stages of PCR analysis.

The PCR laboratory must include the following minimum set of working areas:

• reception, registration, analysis and initial processing of material;
• DNA/RNA isolation;
• preparing reaction mixtures and conducting PCR;
• detection of amplification products by electrophoresis or Hyphae.

It is important to note that when using the method, there is no need to detect amplification products by electrophoresis. Therefore, there is no need to allocate a working area in the laboratory for electrophoresis.

In PCR laboratories, it is also necessary to provide for the presence of auxiliary premises: an archive (for accounting documents), a staff room, a manager’s office, locker rooms for employees, meal rooms, sanitary rooms (toilet), utility rooms (warehouse).

Ideally, it is necessary to have an autoclave room for disinfecting the test material. It may be shared with other departments of the institution, subject to compliance with biological safety requirements.

The premises for performing work at the stages of PCR analysis must be boxed (boxes with pre-boxes). In the area of ​​reception, registration, analysis and primary processing of material carry out material reception, sample preparation (sorting, labeling, centrifugation, etc.), storage and primary inactivation of biomaterial residues with disinfectants. The area for reception, registration, analysis and primary processing of material is located in the material reception room or in a separate boxed room. Here you can also receive and process samples for research using other methods (immunology, for example), provided that a separate equipped workstation is allocated for PCR analysis.

DNA/RNA isolation area placed in a separate room. When organizing a PCR laboratory on the basis of an existing laboratory, DNA/RNA isolation is allowed in premises where other types of research are carried out, except for genetic engineering work. In this case, a working area for DNA/RNA isolation is organized in the room, in which a PCR box or biological safety box is located. No other types of work are allowed in the PCR box for DNA/RNA isolation!

In the area for preparing reaction mixtures and conducting PCR prepare a PCR mixture, add isolated DNA/RNA or cDNA preparations into a PCR tube, reverse transcription of RNA and amplification of DNA or cDNA. The room for preparing reaction mixtures and conducting PCR must be separate. The preparation of PCR reaction mixtures is carried out in a PCR box.

If necessary, the stage of DNA/RNA isolation can be combined in one room with the stage of preparing reaction mixtures and carrying out PCR if there are separate PCR boxes in it - for preparing reaction PCR mixtures and for DNA/RNA isolation.

Detection zone for amplification products located in a separate room, if possible equipped with a PCR box. If it is necessary to simultaneously use the electrophoresis method and the hybridization analysis method to detect amplification products, a separate working area should be allocated in the detection room for hybridization analysis. In this case, equipment and accessories for each type of detection are marked in relation to each zone. Pipettes and glassware intended for electrophoresis are not allowed to be used for hybridization analysis.

Planning decisions and placement of equipment must ensure the flow of movement of the material under study. Air exchange between the amplification product detection room and other rooms should be completely eliminated.

The PCR laboratory is equipped with water supply, sewerage, electricity and heating. All rooms of the PCR laboratory are provided with sufficient natural and artificial lighting.

When constructing new or reconstructing existing PCR laboratories, the premises are equipped with supply and exhaust or exhaust ventilation. The difference in air pressure in the rooms of the PCR laboratory is achieved due to differences in the air exchange rate in them. The air exchange rate must correspond to the values ​​given in the table:

If necessary, air conditioning can be installed in the PCR laboratory.

Interior decoration of premises is carried out in accordance with their functional purpose. The surfaces of walls, floors and ceilings in laboratory premises must be smooth, without cracks, easy to process, and resistant to detergents and disinfectants. Floors should not be slippery. Laboratory furniture must have a coating that is resistant to detergents and disinfectants. The surface of the tables should not have cracks or seams. Laboratory premises must be impenetrable to rodents and insects. The PCR laboratory is provided with fire extinguishing equipment.