ABO Blood Grouping

The principle and the lab guide of ABO blood grouping

1. BLOOD GROUPING M.J. Afra II – BSc Microbiology Department Of Microbiology and Biotechnology Thassim Beevi Abdul Kader College For Women Kilakarai

2. INTRODUCTION There are four main blood groups. They are A, B, AB and O Blood group is determined by the genes you inherit from your parents Each group can be either RhD positive or RhD negative Totally, there are eight main blood groups

3. WHAT IS BLOOD? Blood is a fluid that transport oxygen and nutrients to the cell Blood is made up of Red Blood Cells (RCB), White Blood Cells (WBC) and Platelets The RBC, WBC and Platelets present in the liquid called Plasma . Blood group is identified by antibodies and antigens in the blood

4. ANTIGENS AND ANTIBODIES Antibodies are proteins found in the plasma They are the parts of our body’s natural defenses They recognize the foreign substances and destroy them Antigens are protien molecules that are found on the surface of red blood cells

5. ABO BLOOD GROUPING ABO blood grouping can be performed in two ways: FORWARD GROUPING / CELL GROUPING REVERSE GROUPING / SERUM GROUPING Forward Grouping: Red blood cells ate tested for A and B antigens using known Anti – A and Anti – B sera Reverse Grouping: Serum is tested for Anti – A and Anti – B antibodies using known A and B red cells

6. There are four main blood groups defined by ABO system: Blood Group A – has A antigens on the RBC with Anti – B antibodies in the plasma Blood Group B – has B antigens with Anti – A antibodies in the plasma Blood Group O – has no antigens but both Anti – A and Anti – B antibodies in the plasma Blood Group AB – has both A and B antigens, but no antibodies Blood Group O is the most common blood group. Almost half of the population has blood group O ABO BLOOD GROUPING

7. THE Rh SYSTEM Red blood cells sometimes have another antigen, a protein known as the RhD antigen If RhD antigen is present, your blood group is RhD positive If RhD antigen is absent, your blood group is RhD negative About 85% of the population have Rh positive

8. EIGHT MAIN BLOOD GROUPS A Positive (A+) A Negative (A-) B Positive (B+) B Negative (B-) O Positive (O+) O Negative (O-) AB Positive (AB+) AB Negative (AB-)

9. SAMPLE REQUIRED The blood grouping can be done on whole blood or even on clotted blood The sample can be stored ay 4°C and stable for 5 days Sometimes weak subgroups may result in mistyping where Coombs test may be helpful

10. INDICATIONS ABO blood grouping and Rh typing are done Blood grouping is done for the donor and recipient ( Crossmatch ) Blood grouping is done in the expected mother and new born

11. REAGENTS The blood grouping reagents enables rapid identification of ABO blood group and Rh factor depending upon the antigen present on the surface of red blood cells

12. BLOOD GROUPING REAGENT REAGENTS COLOR VOLUMES STORAGE TEMPERATU-RE Anti A Sera Blue 5 ml 2-8° C Anti B Sera Yellow 5 ml 2-8° C Anti - D Clear 5ml 2-8° C

13. PROCEDURE Inform the patient or individual about the procedure to be carried out Dangle the hand down to increase the flow of blood in the fingers Clean the fingertip to be pierced with spirit or 70% alcohol and gently massage the finger to increase blood flow

14. PROCEDURE 4. With the help of sterile lancet or pricker, pierce the fingertip and place one drop of blood in each of the three cavities 5. Now add one drop of each of the antiserum into each cavities respectively 6. Mix each blood drop with the antiserum using a fresh mixing stick or applicator stick

15. PROCEDURE 7. Now, we can observe the agglutinations in the form of fine red granules within 30 seconds 8. Anti RhD takes slightly longer time to agglutinate compared to Anti A and Anti B

16. EXPECTED RESULTS SLIDE NUMBER ANTI A ANTI B ANTI RhD BLOOD GROUP Slide 1 ✔ ✖ ✔ A+ve Slide 2 ✖ ✔ ✔ B+ve Slide 3 ✔ ✔ ✔ AB+ve Slide 4 ✖ ✖ ✔ O+ve ✔ - Agglutination ✖ - No Agglutination

17. EXPECTED RESULTS SLIDE NUMBER ANTI A ANTI B ANTI RhD BLOOD GROUP Slide 1 ✔ ✖ ✖ A-ve Slide 2 ✖ ✔ ✖ B-ve Slide 3 ✔ ✔ ✖ AB-ve Slide 4 ✖ ✖ ✖ O-ve ✔ - Agglutination ✖ - No Agglutination

18. EXPECTED RESULTS If agglutination is observed when individual’s blood is mixed with Anti A reagent, then the individual is said to have a blood group A If agglutination is observed when individual’s blood is mixed with Anti B reagent, then the individual have a blood group B If agglutination is observed when individual’s blood is mixed with Anti A and Anti B reagent, then the individual is said to have a blood group AB

19. If no agglutination is observed when individual’s blood is mixed with Anti RhD reagent, then the individual is said to have a blood group O If agglutination is observed while mixing the blood with Anti RhD reagent , then the individual is said to have a +ve Rh factor If no agglutination is observed while mixing the blood with Anti RhD reagent , then the individual is said to have –ve Rh factor EXPECTED RESULTS

20. REFERENCE AND SOURCES https://www.labtestsguide.com/blood-group-test-procedure https://www.britannica.com/science/blood-group IMAGE SOURCES http://google.com

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