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Blood grouping

  1. 1. BLOOD GROUPING Dr.Janani Mathialagan, 1st year postgraduate, Pathology
  2. 2. OBJECTIVES – Importance of blood grouping – Landsteiner’s law – Typing techniques – Slide method – Tube method – Cross-matching
  3. 3. NEED FOR BLOOD GROUPING – BLOOD TRANSFUSION – HEMOLYTIC DISEASE OF NEWBORN – PATERNITY DISPUTES – MEDICOLEGAL USE – SUSCEPTIBILITY TO VARIOUS DISEASES – ROUTINE HEALTH CHECKUP
  4. 4. LANDSTEINER’S LAW – If an antigen is present on a patient’s red blood cells, the corresponding antibody will not be present in the patient’s plasma under normal conditions.
  5. 5. Reciprocal relationship between ABO antigens and antibodies Antigens on RBCs Antibody in plasma / serum Blood group A Anti-B A B Anti-A B AB None AB None Anti-A, Anti-B O
  6. 6. ABO antigens & corresponding antibodies
  7. 7. UNIVERSAL DONOR AND RECIPIENT – UNIVERSAL DONOR GROUP O – Neither A or B antigens – UNIVERSAL RECEIPIENT GROUP AB – Patient has no Anti A/Anti B present. – Cannot lyse any transfused cell
  8. 8. ABO TYPING TECHNIQUES – Slide test – Tube technique – Microplate – Gel system
  9. 9. SLIDE GROUPING ADVANTAGES: – Preliminary typing tests – Use during camps DISADVANTAGES: – Not routine test – Less sensitive – Drying of reaction giving to false positive results
  10. 10. ANTISERA
  11. 11. SLIDE GROUPING – Test should be done at room temperature or lower – Tubes, slides should be dry and labeled properly – Antisera should always be added before adding cells – Results should be recorded immediately after observation – Hemolysis is interpreted as positive result
  12. 12. BLOOD SAMPLE
  13. 13. FIRST ADD ANTISERA TO SLIDE ANTI-B ANTI-A ANTI-D
  14. 14. SAMPLES ADDED TO SLIDES ANTI-B ANTI-A ANTI-D
  15. 15. OBSERVE FOR AGGLUTINATION sample 1 ANTI-A ANTI-B ANTI-D
  16. 16. Sample 2 ANTI-A ANTI-B ANTI-D
  17. 17. Test Tube Method Recommended method (Gold standard) – Allows longer incubation of antigen and antibody mixture without drying – Tubes can be centrifuged to enhance reaction – Can detect weaker antigen / antibody Two steps in ABO grouping Cell grouping (Forward grouping) – Tests the patients red cells with known Anti-A & Anti- B to determine the antigen expressed Serum grouping (Reverse grouping) – Test the patients serum with known A & B cells to determine the presence of antibody
  18. 18. CELL GROUPING ( Forward grouping) – Prepare 2-5% suspension of test sample in normal saline – Set three tubes , label them as A,B, D – Add two drops of anti A , anti-B, anti D in three different tubes – Add one drop of 2-5% cell suspension (Ratio of 2:1) – Mix contents well and centrifuge at 1500 rpm for 1 minute – Observe for hemolysis – Gently disperse cell button and check for agglutination – Confirm negative results under microscope
  19. 19. TUBE METHOD
  20. 20. SAMPLE ADDED
  21. 21. MIX TUBE CONTENTS
  22. 22. CENTRIFUGE LOADING
  23. 23. CENTRIFUGE AT 1500 RPM
  24. 24. RH VIEWING BOX
  25. 25. GRADING AGGLUTINATION
  26. 26. SERUM GROUPING ( REVERSE GROUPING) – Prepare 2-5% suspension of pooled cells A,B,O – Label three tubes A cells, B cells and O cells – Place two drops of serum in each tube – Add one drop of cell suspension ( A cell to A tube, B cell to B tube and one drop of O cell to O tube – Centrifuge tubes at 1500 rpm for 1 minute – Gently disperse for agglutination – Negative results check by microscope
  27. 27. 2 vol of test serum/plas ma 1 vol of 5% suspension of reagent red cells in respective tubes Reverse Grouping Centrifuge at 1000 rpm for 1 min Centrifuge & record the results similarly as for cell grouping Shake & leave at room temp (20-24oC) for 5 min
  28. 28. GRADING OF AGGLUTINATION
  29. 29. CROSS MATCHING – A pre-requisite for blood transfusion – Purpose: to avoid reactions of mismatched transfusion
  30. 30. PROCEDURE: – In test tube place 2 drops of recipient’s serum – Add washed donor red cell suspension – Mix and incubate at 37degree C for 30 mins – Centrifuge at 3000 rpm for 1 minute – Examine for agglutination and hemolysis INTERPRETATION: – Matched - no agglutination and hemolysis – Mismatched - either agglutination or hemolysis

Editor's Notes

  • To ensure that serum is added to the test tube it should always be added first to the test tube or slide.
    If red cells are added first, it would be difficult to know if serum was added or not. This may result in wrong results.
  • Take 2 volumes of serum or plasma in a clean labelled test tubes for A cell, B cell and O cells
    Add 5% red cell suspension in respective tubes
    Incubate at room temperature for 5 min
    Centrifuge at 1000 RPM for 1 min
    Look for agglutination

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