Prepare everything you need for blood typing. Technique for determining the blood group of the AB0 system using standard sera. Caring for a central venous catheter

Determination of blood groups by coliclones is carried out in the agglutination reaction. Tsoliclones are saline solutions of monoclonal antibodies for the detection of human erythrocyte antigens. Each reagent is strictly specific to the corresponding antigen. Monoclonal anti-erythrocyte antibodies are produced by mouse hybridomas or human-mouse heterohybridomas.

Analysis methodology

The algorithm for determining the AB0 blood group and the Rh factor by coliclones facilitates and standardizes the typing procedure. Allows unambiguous interpretation of the result of the antigen/antibody reaction. One batch of reagents is sufficient for each study using the AB0 system. The object of the study is whole blood, erythrocyte mass, washed erythrocytes, blood cells in serum or saline. As an example, we give a scheme for typing human blood according to the AB0 antigen system.

Provide bright room lighting and an air temperature of 15 - 25 °C.
Label the wells of the plate: Anti-A, Anti-B, Anti-AB.
Add one drop (approximately 0.1 ml) of Anti-A, Anti-B, Anti-AB reagents to the signed wells.
Place one small drop of concentrated RBC medium (0.01 - 0.03 ml) next to a large drop of each reagent.
In each of the three wells, mix the monoclonal reagent with blood with a separate sterile glass rod.
Gently shake the tablet for 2.5 - 3 minutes. Usually agglutination is formed within 5 - 10 seconds. A margin of time is needed to detect weak variants of antigens.
Perform a visual assessment of the reaction results in each well. The result of the reaction indicates the detection of the corresponding agglutinogen.

Taking a blood sample

Inserting the sample into the well

The tablet with the entered samples

Adding reagents

Tablet with erythrocytes and reagents

Reaction results

Table of results of determining the blood group using coliclones.

«+» - positive result reactions;
"-" - no agglutination.

A negative reaction result in all three wells indicates the absence of agglutinogens. Group: 0(I).
Agglutination with Anti-A and Anti-AB indicates the presence of agglutinogen A. Group affiliation: A(II).
The reaction in the wells with Anti-B and Anti-AB indicates the presence of antigen B. Group affiliation: B(III).
The reaction in all three wells indicates the presence of both antigens. Group: AB(IV).

Mixing of analyzed samples with coliclones

There are two ways to determine blood groups:

using standard sera, when the presence or absence of antigens A and B in erythrocytes is established, i.e., how blood groups are determined in recipients;
using standard sera, antigens are determined in erythrocytes and at the same time, using standard erythrocytes, blood serum (plasma) is examined for the presence of calamus isoagglutinins in it (cross method).

Blood typing equipment

Equipment for determination of blood groups:

2 - physiological solution;

3 - a glass for washing water;

4 - water for washing pipettes and sticks;

7 - five-minute hourglass;

8 - rack with standard sera;

9 - eye spatula for mixing drops;

"Seminars on blood transfusion",

L.V. Ivanov, I.P. Danilov, B.A. Shuvaeva

Please check with your doctor before following any advice.

Rules for determining the blood group by tsoliklons

Very often, blood typing is performed by coliclones. Tsoliklones are divided into anti-A and anti-B. Their use in medicine is popular due to the possibility of determining not only blood types. They also demonstrate Rhesus. For this, the ABO system is used, which replaces conventional sera, the basis of which is the agglutinating isogem.

To determine the group or Rh, the reagents search for the above antigens that are present in red blood cells. For this, antibodies of the standard type are used. Also, agglutins are detected with standard erythrocytes, regardless of whether plasma or special serum is used for research. When it comes to the analysis of donor samples, it is also mandatory to determine the presence of agglutinins in the serum. For this, erythrocytes of a standard type are used.

As a basis for coliclones, hybridoma cell lines are used. They are obtained by combining antibody-forming B-lymphocytes of mouse origin with myeloma cells of the same living organism. With the help of hybridomas, which are individual, antibodies of a homogeneous type are produced. In this case, there can be only one immunoglobulic class as a result. These substances almost completely repeat the structure and activity with biological point vision.

Preparation for work

To determine the blood group and Rhesus with coliclones, there is a special algorithm. This technique of work involves laboratory determination. In the laboratory, certain temperature indicators must be maintained - in the range from 15 to 25 degrees Celsius. In addition, good lighting is required.

All reagents are stored in tightly closed vials. This is necessary to prevent drying out. As a result of this process, the activity of antibodies can be seriously reduced.

For work, it is forbidden to use reagents that are cloudy with the presence of flakes. An individual pipette is used for work. The procedure involves the use of a plate stained in White color well wetted surface. Due to such factors as high avidity and activity of zoliclones, one reagent lot is enough for use.

To perform a blood group test, you first need to prepare all the equipment and reagents. In particular:

a typical plate is being prepared, dry, the entire analysis will be performed on it;
both types of tsoliclones are prepared, as well as two pipettes for working with each separately and two sticks made of glass, which are used for mixing;
you will need a disposable syringe with a needle, which is used to draw blood.

In the sterilized tray, three balls are laid out, which are pre-wetted with alcohol, as well as three sterile wipes. For the fence you will need a rubber band. Collection is carried out in a centrifuge type tube without any liquid. On it with the help of a glassograph, it is required to clearly write the name of the patient. In addition, a form is filled out, which serves as a laboratory referral. The laboratory doctor will re-determine all the main indicators and put a seal and signature.

Definition of results

To get the correct result of the analysis, it is required to carry out an intravenous puncture in compliance with all the rules for sampling. It will be enough to determine five milliliters.

Zoliclones anti-A and anti-B are applied to plates in the laboratory. It will take one large drop. Each is marked with the appropriate inscriptions. Next to them, it is necessary to apply drops of blood in a small amount.

An individual glass rod is used to mix the sample and reagents. The ratio of blood to reagents should be one to ten. Two and a half minutes is enough to observe the reaction.

Just five minutes while stirring the drops is enough to observe the result. It must be evaluated by a doctor. The table below contains the results that you should be guided by when working with serum when using standard type red blood cells.

If the blood group AB is determined in newborns to exclude autoagglutination, then a follow-up study is required. To do this, one drop of isotonic sodium chloride solution is mixed in the above proportion with the blood sent for research. If there is no agglutination reaction, then the blood really corresponds to the AB group.

To determine the Rh factor, the preparation algorithm is the same. After all the items for determining the Rh factor are ready, the study is performed.

This requires a large drop of anti-D-super reagent to be applied to the plate. Nearby, in the same proportion of one to ten, a drop of blood sent for research is applied. Using an individual instrument, the reagent and blood are mixed, then the result is evaluated.

If the agglutination process has begun, then the blood has a positive Rh factor, if the process does not begin, respectively, we are talking about a negative Rh factor. It is worth emphasizing that monoclonal reagents have some advantages over standard sera. In particular, the result in this case is much more accurate, and the procedure itself is safer, painless and does not take much time.

Compilation of kits and determination of group affiliation and Rh factor of blood

Blood typing with anti-A and anti-B coliclones

1. Prepare a plate, soliclones, glass rods, watch, test blood

1. Divide the plate with a felt-tip pen strip into 2 parts

1. Place one drop of anti-A and anti-B zoliclone on a plate

1. Place on a plate one drop of blood (next to a drop of tsoliklon), each of which is 10 times smaller than a drop of tsoliklon

1. Stir with different ends of glass rods

1. Shake for 2.5 minutes

no agglutination - 1 blood group

agglutination in anti-A - 2 blood group

agglutination in anti-B - 3 blood group

agglutination in both drops - 4th blood group

1. Prepare test tubes, pipettes, saline solution (0.9% sodium chloride solution), universal anti-Rhesus reagent, test blood

1. Apply a drop of universal anti-Rhesus reagent and a drop of blood, equal in size, to the test tube wall

1. Shake 3 minutes

1. Add 3 ml. saline

The presence of agglutination - the examined blood is Rh-positive

Absence of agglutination - the tested blood is Rh-negative

Compilation of kits and testing for individual compatibility of donor and recipient blood

ABO compatibility test (by blood group)

1. Prepare a plate, donor blood in a vial, recipient serum in a test tube, watch and glass rods

1. Place a drop of the recipient's serum and a drop of the donor's blood from the vial on the plate

1. Mix with glass rods and shake for 5 minutes

The presence of agglutination - the blood is incompatible for ABO

No agglutination - blood is ABO compatible

Test for compatibility by Rh factor

1. Prepare the donor's blood in a vial, the recipient's serum in a test tube, a test tube, pipettes, polygyukin solution (33%)

1. Into a test tube 2 drops of donor's blood, 1 drop of recipient's blood and 1 drop of polyglucin solution (33%)

1. Shake 5 minutes

1. Add saline solution (2-3 ml)

The presence of agglutination - the blood is incompatible with the Rh factor

Absence of agglutination - the blood is compatible with the Rh factor

Compilation of instrument kits for venesection and catheterization of the subclavian vein

1. Prepare sterile gauze balls in a sterile tray, anatomical tweezers, for processing the surgical field; syringe with novocaine solution for local anesthesia (0.25%); 2 sterile ligatures; 1 sterile catheter and sterile scalpel; sterile needle holder with needle and suture material

1. Ask the patient about the tolerance of novocaine

1. The operation is performed under local anesthesia in the area of the vein of the elbow bend (a vein is removed promptly; 2 ligatures are brought under the vein; the peripheral segment of the vein is tied with the 1st ligature; the vein is incised and a catheter is inserted, which is fixed with a ligature; the wound is sutured; a system is attached to the elastic catheter for blood transfusion)

Set for catheterization of the subclavian vein

1. antiseptic skin treatment, sterile gauze balls on a sterile tray

2. for anesthesia, a syringe with novocaine 0.5% 10 ml

3. disposable subclavian vein catheter

Use of gloves and other personal protective equipment when handling blood

Before working with blood

1. Put on glasses, a mask

1. Treat hands surgically

1. Put on sterile gloves

If the gloves were in contact with the biological media of one patient, then it is unacceptable to touch the skin (wound) of another patient or perform any invasive manipulations on another patient!

After working with blood

1. Immerse gloves in disinfectant solution for 1 hour

1. Place in a special bag (yellow bag, group B)

1. Send for recycling

Holding infusion therapy into the central vein

1. Prepare a sterile tray with a sterile syringe, dressings, a sterile system for the introduction of sterile solutions, two vials of alcohol, tweezers, a tripod, 10% isotonic sodium chloride solution, heparin solution.

2.Fill the drip system for sterile solutions.

3. Collect a sterile syringe, draw up 5 ml of isotonic sodium chloride solution (for washing the catheter).

4. Ask the patient to turn his head in the opposite direction from the subclavian catheter and hold his breath.

5.Remove the plug of the subclavian catheter.

6. Lower the cap into a vial of alcohol.

7. Connect the cannula of a sterile syringe to the subclavian catheter, allow the patient to breathe.

8. Check the presence of the subclavian catheter in the vein (pull the plunger of the syringe towards you), inject 2 ml of isotonic sodium chloride solution when blood appears.

9. Ask the patient to hold his breath.

10. Disconnect the syringe and insert the dropper cannula into the subclavian catheter.

12.Adjust a certain number of drops.

13. Close the lock on the dropper, having finished injecting the sterile solution into the subclavian catheter.

14. Ask the patient to turn his head in the opposite direction from the subclavian catheter and hold his breath.

15. Remove the dropper cannula.

16. Introduce 0.2 ml of heparin with 2 ml of isotonic sodium chloride solution into the subclavian catheter to prevent the formation of blood clots (at the end of the infusion - a heparin lock).

17. Close the entrance to the subclavian catheter with a plug, pulling it out of the vial with alcohol using tweezers. Continue breathing.

NOTE: With prolonged intravenous infusion of sterile solutions, it is necessary to periodically check the presence of the subclavian catheter in the vein under X-ray control.

Caring for a central venous catheter

1. Treat the skin around the catheter with a solution of alcohol (70 degrees), a solution of brilliant green (1%)

1. Check for the absence of kinks, the density of the rubber stopper

1. Check the patency of the catheter: connect a syringe with 20 ml. saline solution, ask the patient to hold his breath while inhaling, pull the piston and the blood should enter the syringe freely

1. Change the infusion system only on inspiration

1. After each infusion, inject 5,000 units of heparin and tightly stopper

44. Application and removal of an interrupted suture Application and removal of an interrupted suture

1) Ask the patient about drug tolerance

2) Prepare anatomical, surgical forceps, needle holder with a needle, silk, scissors on a sterile tray

3) Treat the skin with an antiseptic solution (iodonate) using anatomical tweezers

4) Grab the edge of the wound with surgical tweezers

5) Make an injection, retreating 0.5 cm from the edge of the wound

6) The opposite edge of the wound is stitched opposite the previous one (retreating 0.5 cm)

7) Tie knots on one side of the wound

9) Next seam at a distance of 1 cm

1) Lubricate the wound with iodonate solution

2) Grab the knot with anatomical tweezers and pull towards the scar (not from the scar and not up, because a young, fragile scar can disperse)

3) A white undyed thread appears from the skin; to cross it

Remove thread from seam channel

4) If a drop of blood, pus or lymph appears - inform the doctor, because. it's inflammation

Bandaging of patients with clean and purulent wounds

1. Put on sterile gloves

1. Before removing the bandage dressing (dried to the wound), irrigate the dressing with a solution of hydrogen peroxide (3%)

1. Carefully remove the bandage

1. Wash the wound with a solution of hydrogen peroxide (3%) (has the property of a broad spectrum antiseptic, including destroying clostridia and fungi; eliminates unpleasant odors from the wound and stops bleeding)

1. Apply a napkin moistened with a solution of furacillin (1:5000)

1. Apply a sterile bandage

The procedure is the same. The wound is carried out according to the rule of 3 phases of the wound process: hydration, dehydration and epithelialization.

1. In the hydration phase (there is a lot of pus in the wound and pronounced swelling of the soft tissues)

Dressings are used with water-based ointments (disols with antiseptics), enzymes (trypsin, chymotrypsin, pepsin); physical treatment is contraindicated

2. In the phase of dehydration (little pus, swelling subsides)

Oil-based ointments are used. Dressings can be done less frequently (1 time in 2-3 days). Physiotherapy is possible.

3. In the phase of epithelialization (wound healing is completed)

In general: immunocorrection, detoxification and stimulation of reparative processes in the wound are used.

Exercising toilet wound

The toilet of the wound is carried out with fresh wounds (shallow, in the thickness of the skin, which, at first intention, will not leave a rough scar). In other cases, PST of the wound is performed.

1. Prepare sterile dressing material (napkins, turundas, balls), anatomical tweezers or Billroth clamp, gauze or rubber-gauze drains on a sterile tray

1. Put on sterile gloves

1. Treat the edges of the wound with a solution of alcohol (70 degrees), lubricate with a solution of brilliant green (1%)

1. Apply a sterile bandage or seal with BF-6 glue

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Determination of blood group and Rh factor

Algorithm of actions in determining the blood group and Rh factor

when a woman enters the maternity hospital

Determination of the blood group of the AB0 system by monoclonal antibodies

Prepare:

dry glass slide (typical plate) for determining the blood type;
anti-A (pink) and anti-B (blue) coliclones;
two pipettes for taking tsoliclones from vials;
two glass sticks for mixing the patient's blood with coliclones;
disposable syringe (5-10 ml) with a needle for taking blood from the patient's vein;
put 3 balls moistened with alcohol, 2-3 sterile wipes in a sterile tray;
rubber tourniquet for intravenous punctures;
a dry centrifuge tube, on which the patient's name is clearly signed with a glassgrapher;
form - a referral to the laboratory, where the doctor - laboratory assistant re-determines the blood type, Rh affiliation, stamps and signatures

Observing all the rules for intravenous punctures, take blood from the patient's vein (at least 5 ml).

Zoliclones anti-A and anti-B are applied to the tablet or plate one large drop (0.1) each under the appropriate inscriptions: anti-A and anti-B.
Next to the drops of antibodies, the test blood is applied one small drop (0.01 ml).
After mixing the reagents and blood with different glass rods for anti-A and anti-B in a ratio of 1:10, the agglutination reaction was observed for 2.5 minutes.
Reading the results after 5 minutes while stirring the drops. (from 3 to 5 minutes)
The result is evaluated by a doctor. The evaluation of the results of the agglutination reaction with anti-A and anti-B Zoliclones is presented in the table, which also includes the results of the determination of agglutinins in the serum (plasma) of donors using standard erythrocytes.

In order to exclude autoagglutination, which can be observed in the umbilical cord blood of newborns, if the blood group AB (IV) is established, it is necessary to conduct a control study: mix one drop (0.1 ml) of isotonic sodium chloride solution with a small drop (0.01 ml) of the test blood. The reaction of agglutination should be ABSENT.

A. Carefully fill in all the columns of the form - referral to the laboratory to determine the blood type and Rh affiliation with the signature of the doctor.

C. Deliver the referral form and a tube with the patient's blood to the laboratory for a thorough determination of the patient's blood group and Rh affiliation.

Determination of the Rh factor using a monoclonal reagent (Zoliklon anti-D Super)

A large drop of the reagent (about 0.1 ml) is applied to the plate. A small drop (0.01-0.05 ml) of the test blood is placed nearby and the blood is mixed with the reagent. The agglutination reaction begins to develop in a second, a clearly expressed agglutination occurs in a second. (Rh positive, no agglutination - Rh negative). The results of the reaction are taken into account after 3 minutes.

After mixing the reagent with blood, it is recommended to shake the plate not immediately, but every second, which allows a more complete large-petal agglutination to develop during this time.

Express determination of ABO and Rhesus blood groups using the Erythrotest-Groupcard kit

The set "Erythrotest-Groupcard" (LLC "Hematologist", Moscow) is designed to determine ABO and Rhesus (RhD) human blood groups in laboratory and field conditions. The determination is carried out in a direct hemagglutination reaction and does not require any auxiliary reagents and equipment. The set includes:

card "Erythrotest-Groupcard";
pipette with a dosed drop volume of about 0.02 ml;
sterile packed scarifier;
stick for mixing blood with a reagent.

The wells of the card contain dried anti-A, anti-B, anti-AB and anti-D monoclonal reagents, which immediately dissolve when water is added. Monoclonal anti-D antibodies specifically detect the RhD antigen regardless of the ABO group. The last well contains a solvent for setting a control for non-specific autoagglutination.

The determination is made in native blood with a preservative, in blood without a preservative or blood taken from a finger.

Open the package and take out the card without touching the holes with the working surface. Enter the patient's data, except for the blood group.
Add 1 drop of water (tap or distilled) to each well using the pipette included in the kit. Water is applied to the stain of the dried reagent. Do not allow the drop to dry out.
A small drop of test blood is added to each well. The blood is applied next to the reagent drop, without touching it, to avoid contamination of one reagent with another. A sterile scarifier is used to take blood from a finger.
Thoroughly mix the blood with the reagent using the stick included in the kit. In each well, mixing is done only with a new stick. The use of one mixing stick in different wells is not allowed, as it leads to contamination of the reagents and erroneous determination of the blood group.
Immediately shake the plate. A clear agglutination occurs in seconds, but the final result should be taken into account after 3 minutes, because. in the case of weak forms of antigen A, the reaction occurs later, and the agglutinates are smaller.
The result is evaluated visually (Table 18.6). In the squares under each well enter the result of the reaction with the appropriate reagent (+ or -).

Shelf life of the set "Erythrotest-groupcard" - 2 years at a temperature of 2-8°C. Storage is allowed for 1 year without a refrigerator if the temperature does not exceed 25°C. Storage and use of unsealed or damaged packages is not allowed.

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Technique for determining the blood group of the AB0 system with standard sera

Indications: the need for blood transfusion, preparation for surgery.

Prepare: a standard plate with recesses; a set of glass sticks; isotonic sodium chloride solution; a set of hemagglutinating sera 1, 2, 3, 4 groups of two series; pipettes; blood taken from a vein or finger; watch; trays; gloves; waste material containers; containers with disinfectant solutions.

The nurse is fully prepared to perform the manipulation: dressed in a suit (gown), mask, gloves, cap, removable shoes.
Check the quality of standard hemagglutinating sera by: color marking, appearance(light, transparent); the safety of the packaging, the presence of a properly designed label.
Prepare everything you need to perform the manipulation.

On a white plate, according to the designation, successively apply one drop of serum 1, 2, 3 groups of two series. Each pipette should be immediately lowered into the same ampoule (vial) from which they were taken;

Using a glass rod, apply a drop of the test blood next to the recesses (6 recesses). A drop of blood should be 10 times smaller than a drop of serum;

Mark the time and mix the blood with serum 1 g with a clean, dry glass rod, then 2 g with another stick. etc. in all recesses;

As agglutination occurs, but not earlier than 3 minutes, add one drop of isotonic sodium chloride solution to those drops in which the agglutination reaction occurs to exclude false agglutination and continue monitoring for 5 minutes.

a) with a positive reaction, the smallest grains visible to the eye appear in the mixture, consisting of stuck together erythrocytes. Small grains merge into large grains, and sometimes into flakes, while the whey becomes discolored;

b) when backlash the liquid remains evenly colored pink;

c) 4 combinations of positive and negative reactions are possible:

1. If there is no agglutination in any of the cells, then the blood is group I (0).

2. If agglutination is in the first and third cells, then the blood is II (A) group.

3. If agglutination is in the first and second groups, then the blood of the III (B) group.

4. If agglutination is in the first, second, third cells, then the blood of IV (AB) group.

To eliminate errors, blood is checked with serum of group 4, where there should be no agglutination.

Remove gloves, place them in a disinfectant solution.
Wash hands, dry with a towel.

Note: blood group determination is carried out in a room with good lighting at a temperature of 15 - 25 0 С.

Prepare equipment for determining the blood group and Rh factor;

FEEDING THE PATIENT THROUGH THE PROBE

PREPARE: a sterile thin gastric tube, glycerin, a funnel with a capacity of 200 ml, liquid warm food in the amount of 3-4 glasses (broth, cream, juice), a glass of warm water, a bandage.

1. Invite a doctor.

2. Clear the nasal passages of the patient.

3. Moisten the end of the probe with glycerin, tilt the patient's head slightly forward, help the doctor insert the probe through the nasal passage and esophagus, and then into the stomach.

4. Connect the end of the probe to the funnel.

5. Slowly pour prepared food into the funnel in small portions, and then boiled water.

6. Remove the funnel, rinse.

7. Strengthen the outer end of the probe with a probe inserted through the gastroma, the same food is used, but in small portions (50 ml 8 times a day, gradually increasing to 500 ml per feeding).

1) Examine the edges of the fistulous opening.

2) Make a toilet around the skin of the fistula (treat with alcohol, grease with Lassar paste).

3) Fix the probe with a sticky patch on the skin of the abdomen.

4) Apply a dry sterile bandage.

Equipment for determining the blood type: standard hemagglutinating sera 1,2,3,4 groups of two different series, isotonic sodium chloride solution, pipettes - 7 pieces, glass rods - 7 pieces, scarifier, hourglass, cotton wool, alcohol, plate, rubber gloves , container with disinfectant.

Equipment for determining the Rh - factor: a test tube, isotonic sodium chloride solution, anti-Rh serum, pipettes - 2 pieces, a container with a disinfectant solution.

Order of the FMBA of Russia

dated 30.03.2007 No. 88

FEDERAL MEDICAL AND BIOLOGICAL AGENCY

Compilation of a set and determination of blood grouping according to standard sera

Identification of signs of unsuitability of blood for transfusion.

Indication: determination of the suitability of blood for transfusion.

Equipment: vial or container with blood.

1. Assess the tightness of the package:

The packaging must be absolutely complete;

No traces of integrity violation are unacceptable; if they are present, the blood is unsuitable for transfusion.

2. Assess the correctness of certification:

- the presence of a label with a number;

- designation of blood group and Rh affiliation;

· - surname and initials of the donor;

· - the name of the institution-producer;

· - a stamp about testing for HIV and viral hepatitis.

3. Pay attention to the expiration date of the blood, compare it with the date of transfusion, visually evaluate the blood in the vial:

The blood should be divided into three layers (below - red erythrocytes, above - a narrow gray strip of leukocytes and platelets, above it - yellow transparent plasma);

The plasma must be transparent;

Flakes, films, clots in the plasma indicate its infection and unsuitability for transfusion;

· pink staining plasma indicates hemolysis of red blood cells and the unsuitability of blood for transfusion.

Note: Plasma may be opaque in the case of so-called chylous blood, i.e. blood containing a large amount of neutral fats. When chylous blood is heated to 37 0 C, the plasma becomes transparent, but if the blood is infected, it remains cloudy.

Compilation of a set and determination of blood grouping according to standard sera.

Indications: the need for blood transfusion, preparation for surgery.

· 2 series of standard hemagglutinating sera in special racks;

vial with isotonic sodium chloride solution;

a pipette for taking blood;

pipette for isotonic solution;

hourglass for 5 minutes;

Note. Determination of the blood type is carried out in a room with good lighting and temperature from + 15 0 to +20 0.

Perform manipulation with gloves.

In the presence of damage to the skin, the nurse is temporarily suspended from work.

In case of contact with blood on the skin or mucous membranes, treat according to the current instructions. (see "Asepsis, antisepsis").

1. Check the quality of standard hemagglutinating sera by:

Appearance (light, transparent);

The presence of a correctly designed label indicating the blood type, titer, expiration date, place of preparation.

2.Place on the table:

2 sets of standard hemagglutinating sera of three groups (O, A, B) of two series and one ampoule with AB (IV) serum, each ampoule must have a pipette;

bottle with isotonic solution, pipette;

sterile labeled tablet;

glass slides (glass rods);

a pipette for taking blood;

3. Write on the tablet full name. patient, blood type.

4. Apply one drop (0.1 ml) of standard hemagglutinating sera of three groups of two series to the corresponding wells of the tablet on the tablet.

5. Apply a drop of blood from a finger or from a test tube with a pipette to the appropriate cell.

6. Place one small drop (0.1 ml) of the test blood in each well of the plate, next to the serum, in the ratio of blood: reagent 1:10 (take blood from a large drop using different glass rods).

7. Mix the blood with the reagent, after mixing, gently shake the plate in your hands.

8. Add one drop of 0.9% sodium chloride solution to the drops of serum with red blood cells, where agglutination has occurred, but not earlier than after 3 minutes.

9. Evaluate the result 5 minutes after the start of the reaction:

· - agglutination reaction can be positive and negative;

· - if the sera gave a positive reaction, then the blood contains both AB agglutinogens, in this case, an additional control study should be carried out with standard serum of group AB (IV).

Determination of blood group with standard sera:
It is carried out under the guidance of a doctor.
1. Prepare: plates, pipettes, glass rods, blood vial, balls, alcohol, sera 2 series.
2. Put on sterile clothing.
3. Apply with separate pipettes standard serums according to l-th drop into the cells of a special plate in 2 series (1, 2, 3 groups).
4. Apply a blood smear of each drop of serum with a glass rod.
5. Mix with separate glass rods.
6. Add 1 drop of saline to each series.
7. Observe the reaction for 5 min.
8. Set the reaction with group IV, if agglutination reaction occurred in all 6 cells of the plate, to confirm the result

Determination of the blood group using coliclones:

1. Treat ampoules with zoliclones and ampoules with solvent with alcohol.
2. Open ampoules with anti-A and anti-B coliclones and 2 solvent ampoules
3. Transfer the solvent with separate pipettes to the ampoules with zoliclones.
4. Shake several times, close the ampoules (the obtained reagents can be stored up to 3 months).
5. Apply one large pot of Zoliclon anti-A and anti-B to the cup.
6. Apply one small drop of blood next to a drop of tsoliklon (10 times less).
7. Mix the blood with the Zoliclon solution with separate glass rods.
8. Observe for 2.5 minutes and evaluate the reaction according to the following scheme:

APPENDIX D

Preparing the patient for blood transfusion:

1. 2.

3. 4.

5. 6.

7. 8.

1. Determine the blood type of the patient and the donor.
2. Determine the Rh affiliation of the patient and the donor.
3. Take blood for a general blood test.
4. Take urine for general urinalysis.
5. Check the validity of the blood in the vial.
6. Do not eat 2 hours before a blood transfusion.
7. Empty bladder before blood transfusion.
8. Count the pulse, measure blood pressure and body temperature.
9. Set the devices for individual compatibility by blood group and Rh factor.
10. Place a biological sample.

APPENDIX D

HEMO TRANSFUSION PROTOCOL

(SAMPLE)

1. Patient (name)

2. Case history number __________________

3. Blood type and Rh factor of the patient __________________________

4. Indications for blood transfusion ____________________________________

5. Name of the component ______________________________________

6. Passport data of the component: N __________, donor ________, gr.

blood _______, Rh factor __________, date of collection ___________

7. Name, batch number and amount of resuspension

solution ______________________________

8. Macro-evaluation of the component _______________________________________

9. Results of pre-transfusion study: grouping

patient's blood ___________, component ___________, reaction to

ABO system compatibility ___________

10. Time and method of heating the medium ______________________________

11. Date and time of transfusion __________________________

12. Method and rate of transfusion _________________________________

13. Amount of medium poured __________________________________

14. Patient's condition (pulse, blood pressure, body temperature):

Before transfusion _____________________

During transfusion ___________________

After transfusion:

After 1 hour __________________________

In 2 hours __________________________

After 3 hours __________________________

15. Quantity and macro-evaluation of the first portion of urine __________________

APPENDIX E

QUESTIONNAIRE

I ask you to answer a few questions.

Circle the number of the answer that corresponds to your opinion or write your answer on the free lines. First and last name are not required. The data will only be used in aggregate form.

1. For what indications do you most often transfuse blood? __________________________________________________________

Identification of signs of unsuitability of blood for transfusion.

Indication: determining the suitability of blood for transfusion.

Equipment: vial or container with blood.

Sequencing:

1. Assess the tightness of the package:

The packaging must be absolutely complete;

No traces of integrity violation are unacceptable; if they are present, the blood is unsuitable for transfusion.

2. Assess the correctness of certification:

- the presence of a label with a number;

- dates of procurement;

- designation of blood group and Rh affiliation;

- name of the preservative;

· - surname and initials of the donor;

· - the name of the institution-producer;

- the signature of the doctor;

· - a stamp about testing for HIV and viral hepatitis.

3. Pay attention to the expiration date of the blood, compare it with the date of transfusion, visually evaluate the blood in the vial:

The blood should be divided into three layers (below - red erythrocytes, above - a narrow gray strip of leukocytes and platelets, above it - yellow transparent plasma);

The plasma must be transparent;

Flakes, films, clots in the plasma indicate its infection and unsuitability for transfusion;

Pink coloration of plasma indicates hemolysis of erythrocytes and the unsuitability of blood for transfusion.

Note: Plasma may be opaque in the case of so-called chylous blood, i.e. blood containing a large amount of neutral fats. When chylous blood is heated to 37 0 C, the plasma becomes transparent, but if the blood is infected, it remains cloudy.

Compilation of a set and determination of blood grouping according to standard sera.

Indications: the need for blood transfusion, preparation for surgery.

Equipment:

· 2 series of standard hemagglutinating sera in special racks;

vial with isotonic sodium chloride solution;

Marked tablets

a pipette for taking blood;

pipette for isotonic solution;

hourglass for 5 minutes;

· gloves.

Note. Determination of the blood type is carried out in a room with good lighting and temperature from + 15 0 to +20 0.

Perform manipulation with gloves.

In the presence of damage to the skin, the nurse is temporarily suspended from work.

In case of contact with blood on the skin or mucous membranes, treat according to the current instructions. (see "Asepsis, antisepsis").

Sequencing:

1. Check the quality of standard hemagglutinating sera by:

color marking;

Appearance (light, transparent);

preservation of the ampoule;

The presence of a correctly designed label indicating the blood type, titer, expiration date, place of preparation.

2.Place on the table:

2 sets of standard hemagglutinating sera of three groups (O, A, B) of two series and one ampoule with AB (IV) serum, each ampoule must have a pipette;

bottle with isotonic solution, pipette;

sterile labeled tablet;

glass slides (glass rods);

a pipette for taking blood;

· hourglass.

3. Write on the tablet full name. patient, blood type.

4. Apply one drop (0.1 ml) of standard hemagglutinating sera of three groups of two series to the corresponding wells of the tablet on the tablet.

5. Apply a drop of blood from a finger or from a test tube with a pipette to the appropriate cell.

6. Place one small drop (0.1 ml) of the test blood in each well of the plate, next to the serum, in the ratio of blood: reagent 1:10 (take blood from a large drop using different glass rods).

7. Mix the blood with the reagent, after mixing, gently shake the plate in your hands.

8. Add one drop of 0.9% sodium chloride solution to the drops of serum with red blood cells, where agglutination has occurred, but not earlier than after 3 minutes.

9. Evaluate the result 5 minutes after the start of the reaction:

· - agglutination reaction can be positive and negative;

· - if the sera gave a positive reaction, then the blood contains both AB agglutinogens, in this case, an additional control study should be carried out with standard serum of group AB (IV).

Related information:

III. Measurement technique and calculation formulas. I. Objectives of the work: determination of the acceleration of free fall by the period of oscillation of the mathematical and reversible physical pendulums

IN modern medicine blood group characterizes a set of antigens located on the surface of erythrocytes, which determine their specificity. There are a huge number of such antigens (usually a table of blood groups with various antigens is used), but the determination of the blood group is generally performed using the classification according to the Rh factor and the AB0 system.

Defining a group is a mandatory procedure in preparation for any operation. Such an analysis is also necessary when entering the service in some contingents, including the military, employees of internal organs and law enforcement agencies. This intervention is carried out due to the increased risk of a life-threatening condition in order to reduce the time required to provide assistance in the form of a blood transfusion.

The composition of the blood of various blood groups

The essence of the AB0 system is the presence of antigen structures on erythrocytes. In plasma, there are no typical antibodies corresponding to them (gamma globulins). Therefore, for a blood test, you can use the "antigen + antibody" reaction.

Red blood cells stick together when antigen and antibody meet. This reaction is called hemagglutination. The reaction appears as small flakes during analysis. The study is based on sera agglutination imaging.

The erythrocyte antigens "A" bind to antibodies "ά", as well as "B" to "β", respectively.

The following blood groups are distinguished by composition:

I (0) - ά, β - the surface of erythrocytes does not contain antigens at all;
II (A) - β - on the surface there is antigen A and antibody β;
III (B) – ά - surface contains B with ά-type antibody;
IV (AB) - 00 - the surface contains both antigens, but does not have antibodies.

The embryo already has antigens in the state of the embryo, and agglutinins (antibodies) appear in the first month of life.

Methods of determination

Standard Method

There are many techniques, but the laboratory usually uses standard sera.

The standard sera method is used to determine the types of AB0 antigens. The composition of the standard isohemagglutinating serum contains a set of antibodies to erythrocyte molecules. In the presence of an antigen that is susceptible to the action of antibodies, an antigen-antibody complex is formed, which triggers a cascade of immune responses.

The result of this reaction is the agglutination of erythrocytes, based on the nature of the ongoing agglutination, it is possible to determine whether the sample belongs to any group.

For the preparation of standard serum, donor blood is used and a certain system is used - by isolating plasma, including antibodies, and then diluting it. Dilution is performed using isotonic sodium chloride solution.

Breeding is done like this:

The study itself is carried out in the following way:

A drop of each serum (with a total volume of approximately 0.1 milliliter) is placed on a special tablet on the area where there is a corresponding mark (2 samples are used, one of them is control, the second is intended for research).
Then, next to each drop of serum, the test sample is placed in a volume of 0.01 milliliter, after which it is separately mixed with each diagnosticum.

Rules for decoding results

After five minutes, you can evaluate the results of the study. In large drops of serum, clarification occurs, in some there is an agglutination reaction (small flakes are formed), in others it is not.

Video: Determination of blood group and Rh factor

Here are the possible options:

If there is no agglutination reaction in both samples with sera II and III (+ control 1 and IV) - the definition of the first group;
If clotting is observed in all samples except II - the definition of the second;
In the absence of an agglutination reaction only in a sample from group III - definition III;
If clotting is observed in all samples, including the IV-control - the definition of IV.

When the sera are arranged in the correct order and labels are on the plate, it is easy to navigate: the group corresponds to places with no agglutination.

In some cases, bonding is not clearly visible. Then the analysis needs to be redone, fine agglutination is observed under a microscope.

Cross reaction method

The essence of this technique is the determination of agglutinogens using standard sera or coliclones with a parallel determination of agglutinins using reference erythrocytes.

The technique of analysis by the cross method has practically no differences from the study using sera, but there are some additions.

Drop by drop of standard erythrocytes should be added to the plate under the sera. Then, plasma is removed from the tube with the patient's blood, which has passed through the centrifuge, with a pipette, which is placed on the standard erythrocytes at the bottom - added to the standard serum.

As well as according to the technique of the standard method, the results of the study are evaluated a few minutes after the start of the reaction. In the case of an agglutination reaction, we can talk about the presence of AB0 agglutinins, in the case of a plasma reaction, we can judge about agglutinogens.

The results of a blood test using standard erythrocytes and sera:

The presence of agglutination in the reaction with standard isohemagglutinating sera				The presence of agglutination in the reaction with standard erythrocytes			Blood groups
0(I)	A(II)	B(III)	AB(IV)	0(I)	A(II)	B(III)
–	–	–	-	–	+	+	0(I)
+	–	+	-	–	–	+	A(II)
+	+	–	-	–	+	–	B(III)
+	+	+	–	–	–	–	AB(IV)

Agglutination;

- no agglutination;

- no reaction is carried out.

The cross method has become widespread due to the fact that it prevents diagnostic errors that occur when using standard methods.

Determination of the blood group by tsoliklons

Tsoliklons are synthetic serum substitutes that contain artificial substitutes for ά and β agglutinins. They are called erythrotests "Tsoliklon anti-A" (have a pink color), as well as "anti-B" (have Blue colour). The expected agglutination is observed between coliclon agglutinins and blood erythrocytes.

This technique does not require two series, it is more reliable and accurate. Conducting a study and evaluating its results occur in the same way as in the standard method.

	Type of tsoliklon		Blood type
Result of agglutination	Anti-A	Anti-B
	-	-	0(I)
	+	-	A(II)
	-	+	B(III)
	+	+	AB(IV)

Group IV (AB) is necessarily confirmed by agglutination with anti-AB coliclone, as well as by the absence of erythrocyte agglutination in isotonic sodium chloride solution.

Express method using the set "Erythrotest-Groupcard"

Although the generally accepted methods for determining blood belonging to a particular group are widespread, in modern medicine there is an introduction of express methods, the most common of which is "Erythrotest".

When determining a group using the “Erythrotest groupcard” method, a set of tools is required, including the following devices:

A tablet with five wells to determine the group by its Rh-affiliation and the AB0 system;
Scarifier designed to obtain a sample required for research;
Glass rods for sample mixing;
A clean pipette for a set of solutions.

All of these tools are necessary for error-free diagnostics.

The Erythrotest-Groupcard blood test kit allows you to study the Rh factor and determine the blood group in any conditions, it is especially effective in the absence of the possibility of using conventional methods.

In the wells on the plate there are tsoliclones to antigens (these are anti-A, -B, -AB tsoliklones) and to the main antigen, which determines the inheritance of the Rh factor (it is tsoliklon anti-D). The fifth well contains a control reagent, which allows you to prevent possible errors and correctly determine belonging to a blood group.

Prepare everything you need for blood typing. Technique for determining the blood group of the AB0 system using standard sera. Caring for a central venous catheter

Analysis methodology

Reaction results

Rules for determining the blood group by tsoliklons

Preparation for work

Definition of results

Compilation of kits and determination of group affiliation and Rh factor of blood

Blood typing equipment

Technique for determining the blood group of the AB0 system with standard sera

Prepare equipment for determining the blood group and Rh factor;

Compilation of a set and determination of blood grouping according to standard sera

The composition of the blood of various blood groups

Methods of determination

Standard Method

Rules for decoding results

Video: Determination of blood group and Rh factor

Cross reaction method

Determination of the blood group by tsoliklons

Express method using the set "Erythrotest-Groupcard"

Video: Determination of blood groups using tsoliklon