University:
University of Cincinnati Blue Ash
Course:
BIOL 5180 | Immunology
Academic year:

2024
Views:

344

Pages:

3
Author:

Jasper J.

ABO SYSTEM BLOOD GROUP EXAMINATION(TUBE) Principle : Antigen + Antibody ------------ Agglutination Objective : To determine the presence / absence of antigens on cellsred blood (cell grouping) and to determine the presence/absence of antibodies in serum/plasma (serum grouping) Utility : To determine a person's blood type Method : Tube Test Material : Blood sample to be examined Reagents : Leave the reagent at room temperature & note the lot.no as wellexp. date 1. Anti-A Sera Test 2. Anti B Sera Test 3. Cell A Test 10 4. B Cell Test 10% 5. O Cell Test 10% 6. Anti-D Sera Test 7. Bovine Albumin 6% 8. Saline Tool : 1. Centrifuge 2. Reaction Tubes 3. 1 ml plastic pipette 4. Tube Rack 5. Object Glass 6. Waste Container 7. Microscope 8. Rinsing Glass/Becker Glass Ways of working : 1. Prepare 8 tubes on a rack ● Label tube 1: -A ● Label tube 2: -B ● Label tube 3: EA ● Label tube 4: EB ● Label tube 5: EO ● Label tube 6: AK ● Label tube 7: -D ● Label tube 8: BA 2. Fill each tube with: ●Tube 1 : 2 drops anti A ●Tube 2 : 2 drops anti B ●Tube 3 : 1 drop of A cell test 5% ●Tube 4 : 1 drop of B cell test 5% ●Tube 5 : : 1 drop of O cell test 5% ●Tube 6 : see number 3 & 4 ●Tube 7 : 2 drops of anti-D ●Tube 8 : 2 drops bovine albumin 6% 3. Place 1 drop each of the red blood cells to be examined in 5% suspension in tubes 1, 2, 6, 7. And 8 4. Drop 2 drops each of serum/plasma to be examined in tubes 3, 4, 5 and 6 5. Shake all the tubes until evenly mixed 6. Rotate at 3000 rpm for 15 seconds or incubate for 60 minutes at room temperature Reading Results: 1. Read the reaction by shaking the tube slowly while paying attention to the degree of agglutination 2. If the red blood cells being examined occur: ● Agglutination : There are antigens on red blood cells ● No agglutination occurs : There are no antigens on red blood cells 3. If the serum/plasma being examined occurs: ● Agglutination : There are antibodies in the serum/plasma ● No agglutination occurs : There are no antibodies in the serum/plasma 4. Determine the degree of agglutination: ● + + + + (4+ ) : Large clots with clear fluid around them ● + + + (3+ ) : : Several large lumps with clear fluid around them ● + + (2+ ) : Several rather large lumps with slightly red fluid surrounding ● + (1+ ) : Small clots with slightly red fluid around them ● ± ( 1W ) : Clots that are not clearly visible require help microscope ● Lysis : Red blood cellls are clear red ● - /0 (Negative) : Suspended/homogeneous CONCLUSION RESULT In the ABO Blood Group System: 1. If agglutination occurs in the Anti-A and B cell tests, the blood type being tested is A 2. If there is agglutination in the Anti B and A cell tests, the blood type being tested is B 3. If agglutination occurs in Anti A and Anti B, and there is no agglutination in the A cell test and B cell test, then the blood type examined is AB 4. If there is no agglutination in the Anti A, Anti B and there is agglutination in the A cell test and B cell test, then the blood type examined is O 5. Sec O and Auto Control tests must be negative In Rhesus Factor: 6. If there is agglutination of anti-D then the blood type examined is Rhesus Positive (Rh.D+) 7. If there is no agglutination of anti-D then the blood type being examined is Rhesus Negative (Rh.Negatif)