STANDARD OPERATING PROCEDURE: TO ENSURE RELIABILITY AND REPRODUCIBILITY OF BLOOD GROUP RESULTS

BLOOD CENTRE NAME

SOP Number	Effective date	Pages	Author	Authorised by
Version	Review Period	No. of copies	Reviewed by	Date

LOCATION     Red Cell Serology Laboratory	SUBJECT       To ensure reliability and reproducibility of blood group results
FUNCTION          Daily Quality Control of ABO & Rh Blood Group reagents	DISTRIBUTION – Supervisor in charge Red Cell Serology Laboratory – Master File

1. SCOPE & APPLICATION

This Standard Operating Procedure (SOP) provides the daily checks on blood group reagents to ensure reliability and reproducibility of blood group results.

2. RESPONSIBILITY

It is the responsibility of the technician / supervisor in the red cell serology laboratory to ensure that quality controlled reagents and proper cell concentrations are used for testing for which daily quality control checks and test controls are used with proper documentation. The reagents should be stored and used as per manufacturer’s instruction.

Any fault in the reagents should be immediately reported to the Quality Assurance Manager.

• REFERENCES

1. Technical Manual of the American Association of Blood Banks 20^th edition 2020

• MATERIALS  REQUIRED

4.1       Equipment:

• Refrigerator to store samples and reagents at 2-6⁰C.

• Table top Centrifuge.

• Automated Cell Washer.

• Microscope.

4.2       Reagents:

• Anti-A, Anti-B, Anti-AB, Anti  D(Monoclonal and Bioclone) Antisera.

• Clotted or anticoagulated blood samples of random blood donors.

• Group A,B and O pooled Cells.

• 0.9%  saline.

4.3      Glassware:

• Serum tubes.

• Micro tubes.

• Pasteur pipettes.

• Glass slides.

4.4       Miscellaneous:

• Rubber teats.

• Disposal box.

• 2 plastic beakers.

• Wooden block.

• Aluminium racks.

• PROCEDURE

5.1      Principle:

Test for reactivity and specificity is based on the principle of agglutination of antigen positive red cells in the presence of antibody directed towards the antigen.

5.2       Quality Control Checks:

5.2.1     Visual Inspection:

Examine each vial carefully for precipitate, gel formation, turbidity or change in colour.

5.2.2     Reactivity and Specificity:

1. Add one drop of 3 5% suspension of the appropriate red cells to the one-drop of antiserum in a microtube.

• Mix well and incubate (as per manufacturer’s instruction).

3.   Note the reactions as  under:

	RED CELLS FOR TESTING
	POSITIVE REACTORS	NEGATIVE REACTORS
Anti-A	Pooled A Cells	Pooled B, Pooled O Cells
Anti-B	Pooled B Cells	Pooled A, Pooled O Cells
Anti-AB	Pooled A, Pooled B Cells	Pooled O Cells
Anti-D Bioclone	RhD- positive cells	RhD- negative cells
	(any ABO group)	(any ABO group)
Anti-D Monoclonal	RhD- positive	RhD- negative
	(any ABO group)	(any ABO group)

5.3      Results:

5.3.1     Visual Inspection:

• Record presence or absence of precipitate, gel formation, turbidity or colour change.

5.3.2     Reactivity and Specificity:

• Centrifuge the Tubes (as per manufacturer’s instruction).

• Resuspend the red cell button and examine for agglutination / haemolysis.

• Grade and record test results.

5.4      Interpretation:

5.4.1     Visual Inspection:

• The presence of precipitate, gel formation, turbidity, colour change indicates that the reagent is contaminated and should not be used.

• The absence of all the above indicates that the reagent is ‘clear’ and suitable for use.

5.4.2     Reactivity and Specificity:

• Agglutination of specific red cells is a positive reaction and indicates the reactivity of the corresponding antibody in the reagent. The expected agglutination reaction for positive test is +3 to +4.

• The absence of agglutination / haemolysis is considered to be a negative reaction and indicates the absence of the corresponding antibody specificity in the reagent.

• Clear cut negative reactions with the negative reactors rules out the presence of irregular agglutinins and haemolysis in the reagent.

6. DOCUMENTATION

Enter    the    results      in   the    Blood    Group    Register      in   the    Red    Cell    Serology

Laboratory. Enter identification number of the individual donor cells used for pooling and the reaction strengths. Sign the results as the individual preparing the pooled cells and testing the reagent.

7. END OF DOCUMENT