Crossmatching
- 1. CROSS-MATCHING DR.MD.MIZANUR RAHMAN CHOWDHURY SPECIALIST TRANSFUSION MEDICINE UNITED HOSPITAL LTD
- 2. INTRODUCTION Cross-matching is one of the most important serological procedure pertaining to blood group serology and is the fundamental procedure responsible for safe blood transfusion. Basically Cross-matching is an antigen-antibody reaction, a correct interpretation of which is the most essential preliminary step in the practice of safe transfusion of blood. By cross-matching we are able to detect the atypical and clinically significant antibody mostly IgM and IgG present in recipient serum or in donor serum, also by autocontrol we are able to detect auto-antibody in patient himself.
- 3. CONTINUE… Cross match test is carried out to ensure that there are no antibodies present in patients serum that will react with donor cells when transfused. Unless there is an urgent need for blood, a cross-match must be preformed for red cell transfusion.
- 4. FUNCTIONS OF CROSS-MATCH It is final check of ABO compatibility between the donor and patient. It may detect the presence of an antibody in the patient’s serum which will react with an antigen on donor red cells To ensure that patient/ recipient is supplied with a compitable unit of antigen negative blood. To prevent hemolytic transfusion reaction. To detect immunologic auto antigen and auto antibody, and blood to be issued has to be processed accordingly. All this done with sole aim for safe transfusion of blood
- 5. CROSS MATCH TESTING PROCEDURES HAD BEEN DIVIDED INTO TWO PARTS….. Major cross match Minor cross
- 6. MAJOR CROSS MATCH DONOR RED CELLS + PATIENT SERUM
- 7. MINOR CROSS MATCH PATIENT CELLS + DONOR SERUM
- 8. PRINCIPLE OF CROSS MATCH Major cross match is done to detect any serological incompatibility b/w donor’s cells and patients serum. Minor cross-match is done to detect any serological incompatibility b/w patient cells and donor serum. Cross-match is verified with a coombs reaction to detect even the incomplete antibodies.
- 9. CROSS-MATCH TECHNIQUES Immediate spin method Saline room temperature technique Indirect Antiglobulin technique Albumin addition technique
- 10. IMMEDIATE SPIN TECHNIQUE/SALINE ROOM TEMPERATURE TECHNIQUE Immediate spin technique or saline room temperature technique is enough to rule out any ABO grouping error. But this technique is inadequate for identification of clinically significant IgG type of antibodies. Both these techniques are not good, because antibody screening has not been carried out in our country.
- 11. PROCEDURE label the tube as major/minor cross-match with donor number & patient ID. Using a micropipette add 1 volume of 2-5 % red cell suspension to the labeled tube and 2 volume of serum to the same tube. Mix the tube well and centrifuge at 1000RPM for 1 min.
- 12. CONTINUE.. Gently shake and observe for agglutination or hemolysis NO agglutination or hemolysis: Compatible Agglutination or hemolysis seen : Incompatible
- 14. INDIRECT ANTI GLOBULIN TECHNIQUE (IAT) IAT test is widely used in cross matching as it detects majority of incomplete antibodies.
- 15. PROCEDURE Put 2 drops of serum in prelabelled test tube, & one drop of 2 – 5% suspension of red cells Incubate 45 to 60 min at 37°c Gently shake and observe for agglutination or hemolysis If test tube show any agglutination or hemolysis that means incompatible at 37°C
- 16. CONTINUE… If no agglutination or hemolysis present, wash the cells three to four times with saline and decant the last wash completely. Add 2 drops of AHG reagent to the test tube Gently shake and read the result immediately, if no reaction, Wait for 5 min Centrifuge at 1000rpm for 1 min
- 17. CONTINUE… Shake gently and observe for agglutination or hemolysis with optical aid NO agglutination or hemolysis: Compatible Agglutination or hemolysis seen : Incompatible
- 18. CONTINUE.. If the test is negative add one drop control IgG coated red cells. Centrifuge again at 1000 rpm for 1 min look for hemolysis or agglutination if no agglutination or hemolysis, the test is invalid repeat the procedure
- 19. INTERPRETATION Haemolysis or agglutination at any stage of the test procedure except after adding control IgG coated red cells indicates incompatible
- 20. Indirect Anti globulin Technique
- 21. FACTORS AFFECTING IAT Temperature ( 37°C) Serum : Cell ratio Incubation time Suspending Medium ( sensitivity of IAT increased with addition of 22 % bovine albumin , enzyme or LISS)
- 22. ALBUMIN ADDITION TECHNIQUE Put 2drops of serum of serum 1drop of 2-5% red cell suspension in to a pre labeled test tube, Gently shake the tube Add 2 drops of bovine serum albumin & Gently shake the tube Incubate the test tube 30min at 37°c Centrifuge 1000rpm for 1 min
- 23. CONTINUE… Wash 3 times(minute amount of human protein can neutralize AHG )ensure that the saline is completely decanted after each wash Add AHG 2 drops to the dry cell button Gently shake and read the result immediately, if no reaction, Wait for 5 min Centrifuge at 1000rpm for 1 min
- 24. CONTINUE… Shake gently and observe for agglutination or hemolysis with optical aid NO agglutination or hemolysis: Compatible Agglutination or hemolysis seen : Incompatible
- 25. CONTINUE.. If the test is negative add one drop control IgG coated red cells. Centrifuge again at 1000 rpm for 1 min look for hemolysis or agglutination if no agglutination or hemolysis, the test is invalid repeat the procedure
- 26. 2 drops BSA Albumin Addition Technique
- 28. CAUSES OF POSITIVE RESULTS IN CROSS-MATCH Incorrect ABO grouping of patient or donor. An allo-antibody in the patient serum reacting with the corresponding antigens on donor cells. An auto-antibody in patients serum reacting with corresponding antigen on donor red cells. This can be solved by putting auto control which will be positive. Donor red cells with a positive DAT. Problems in patients serum E.g. Multiple myeloma. Dirty glass ware.
- 29. DRAWBACK IN CROSS-MATCHING Rh typing errors cannot be detected by cross-matching. It can detect only antibody specific for red cell antigen of the donor. A donor unit without appropriate antigen or with very weak antigen may thus fail to detect the corresponding antibody even if it present in serum of the patient Present method of compatibility testing cannot detect any antibodies to either leucocyte antigen or platelet antigen
- 30. Thank you