Republic of the Philippines MS-PAT-WI-

Province of Sultan Kudarat

Document No. BB-02
SULTAN KUDARAT PROVINCIAL HOSPITAL
Revision No. 1
LABORATORY DEPARTMENT
BLOOD BANK OF OPERATIONS Effectivity Date JULY 16, 2024

ABO REVERSE GROUPING Page No. 4 OF 4

I. POLICY
All Medical Technologists rotating in Blood Bank Section shall be oriented, guided
and knowledgeable in performing the ABO reverse grouping using the tube method
and grading of agglutintion of Known A and Known B cells.

II. PURPOSE
To establish a documented work instruction for ABO reverse grouping that uses the
patient's plasma or serum, combined with reagent group A and group B red blood
cells, to determine which ABO antibodies are present. This doucment will serve as
protocol on how to determine blood type through ABO reverse grouping.

III. SCOPE
This document covers the procedure of blood typing with the use of known red cell
samples against pateint’s serum sample.

IV. RESPONSIBILITIES

Laboratory Personnel - responsible for request and specimen reception, request and
specimen forwarding, endorsing or releasing of result.

Medical Technologist - responsible for request and specimen receiving, processing,

recording of result, encoding and printing of result, and specimen management.

Quality assurance officer- responsible in validating the results released by MedTech.

V. DEFINITION OF TERMS

ABO blood group - the classification of human blood based on the inherited properties
of red blood cells (erythrocytes) as determined by the presence or absence of the
antigens A and B, which are carried on the surface of the red cells. Persons may thus
have type A, type B, type O, or type AB blood.

Agglutination - is an antigen-antibody reaction that occurs when an antigen (i.e., a

molecule capable of triggering the adaptive immune response) is mixed with its
corresponding antibody at a suitable pH and temperature

Prepared by: Reviewed by: Approved by: Noted by:

CHARINA E. GENOTIVA, SHERJAN P. KALIM, MD, REX ARCHANGEL S. LAMPREA,

CHARMA GRACE E. FPSP, MHcA
RMT, MPA MD, FPSMS
FRIAS, RMT
BLOOD BANK SECTION HEAD Chief Medical Technologist Head of the Laboratory OIC-Chief of Hospital
Date: July 16, 2024 Date: July 16, 2024 Date: July 16, 2024 Date: July 16, 2024
 Republic of the Philippines MS-PAT-WI-
Province of Sultan Kudarat
Document No. BB-02
SULTAN KUDARAT PROVINCIAL HOSPITAL
Revision No. 1
LABORATORY DEPARTMENT
BLOOD BANK OF OPERATIONS Effectivity Date JULY 16, 2024

ABO REVERSE GROUPING Page No. 4 OF 4

VI. PROCEDURE
6.1. SAMPLE PROCESSING PROCEDURE
6.1.1. Principle

6.1.1.1. The reverse blood grouping procedure is based on the principle of direct
hemagglutination. The erythrocytes of a person contain blood group antigens on
the surface of the membrane. When these antigens are allowed to treat with
corresponding antibodies, antigen-antibody reaction occurs and form
agglutination.

6.1.2. Sample

6.1.2.1. Serum Sample

6.1.3. Reagents and supplies

6.1.3.1. Test tube rack

6.1.3.2. Test tubes
6.1.3.3. Centrifuge
6.1.3.4. Serological Pipette
6.1.3.5. Normal Saline Solution
6.1.3.6. Anticoagulated blood preferably with EDTA
6.1.3.7. Parafilm
6.1.3.8. Known A, B, AB and O 2-5% red cell suspension
6.1.3.9. Normal Saline Solution (0.85-0.9% NaCl) in wash bottle

VII. WORK INSTRUCTION

Prepared by: Reviewed by: Approved by: Noted by:

CHARINA E. GENOTIVA, SHERJAN P. KALIM, MD, REX ARCHANGEL S. LAMPREA,

CHARMA GRACE E. FPSP, MHcA
RMT, MPA MD, FPSMS
FRIAS, RMT
BLOOD BANK SECTION HEAD Chief Medical Technologist Head of the Laboratory OIC-Chief of Hospital
Date: July 16, 2024 Date: July 16, 2024 Date: July 16, 2024 Date: July 16, 2024
 Republic of the Philippines MS-PAT-WI-
Province of Sultan Kudarat
Document No. BB-02
SULTAN KUDARAT PROVINCIAL HOSPITAL
Revision No. 1
LABORATORY DEPARTMENT
BLOOD BANK OF OPERATIONS Effectivity Date JULY 16, 2024

ABO REVERSE GROUPING Page No. 4 OF 4

The Medical Technologist shall:

7.1. Prepare 2-5% red cell suspension of A, B, AB and O cells

7.2. Label the test tubes as A, B, AB and O respectively
7.3. Follow the table below.

Content A B AB O

Unknown Serum 2 drops 2 drops 2 drops 2 drops

A-cell 1 drop ---- --- ---

B-cell --- 1 drop --- ---

AB-cell --- --- 1 drop ---

O-cell --- --- --- 1 drop

7.4. Cover the tubes with parafilm.

7.4. Mix gently and centrifuge for 15 seconds at 3,400 rpm
7.5. Gently dislodge the cell button and examine for agglutination or hemolysis
7.6. Grade each reaction, encode and print result.

Prepared by: Reviewed by: Approved by: Noted by:

CHARINA E. GENOTIVA, SHERJAN P. KALIM, MD, REX ARCHANGEL S. LAMPREA,

CHARMA GRACE E. FPSP, MHcA
RMT, MPA MD, FPSMS
FRIAS, RMT
BLOOD BANK SECTION HEAD Chief Medical Technologist Head of the Laboratory OIC-Chief of Hospital
Date: July 16, 2024 Date: July 16, 2024 Date: July 16, 2024 Date: July 16, 2024
 Republic of the Philippines MS-PAT-WI-
Province of Sultan Kudarat
Document No. BB-02
SULTAN KUDARAT PROVINCIAL HOSPITAL
Revision No. 1
LABORATORY DEPARTMENT
BLOOD BANK OF OPERATIONS Effectivity Date JULY 16, 2024

ABO REVERSE GROUPING Page No. 4 OF 4

7.7. Verify result by the Quality Assurance Officer or Senior Medicla Technologist.
7.8. Record the result to the blood typing result loogbook.
7.9. Charge the laboratory request to the patient’s IHOMIS account.
7.10. Release result to the receptionist.

Guide in the interpreation of Forward Blood Grouping

Content Known A-cells Known B-cells Known AB-cells Known O-cells

A o + + o

B + o + o

AB o o o o

O + + + o

[+] with agglutination [o] no agglutination

 Agglutination in B and AB cells and no agglutination in A cells demonstrates the presence

of Anti-B in the serum, thus, indicating that the individual is type A.
 Agglutination in A and AB cells and no agglutination in B cells demonstrates the presence
of Anti-A in the serum, thus, indicating that the individual is type B.

Prepared by: Reviewed by: Approved by: Noted by:

CHARINA E. GENOTIVA, SHERJAN P. KALIM, MD, REX ARCHANGEL S. LAMPREA,

CHARMA GRACE E. FPSP, MHcA
RMT, MPA MD, FPSMS
FRIAS, RMT
BLOOD BANK SECTION HEAD Chief Medical Technologist Head of the Laboratory OIC-Chief of Hospital
Date: July 16, 2024 Date: July 16, 2024 Date: July 16, 2024 Date: July 16, 2024
 Republic of the Philippines MS-PAT-WI-
Province of Sultan Kudarat
Document No. BB-02
SULTAN KUDARAT PROVINCIAL HOSPITAL
Revision No. 1
LABORATORY DEPARTMENT
BLOOD BANK OF OPERATIONS Effectivity Date JULY 16, 2024

ABO REVERSE GROUPING Page No. 4 OF 4

 Agglutination in A,B and AB cells demonstrates the presence of Anti-A and Anti-B in the
serum, thus, indicating that the individual is type O.
 No agglutination in tubes A, B, and AB cells, demonstrates the absence of Anti-A and Anti-
B in the serum, thus indicating that the individual is type AB.

Prepared by: Reviewed by: Approved by: Noted by:

CHARINA E. GENOTIVA, SHERJAN P. KALIM, MD, REX ARCHANGEL S. LAMPREA,

CHARMA GRACE E. FPSP, MHcA
RMT, MPA MD, FPSMS
FRIAS, RMT
BLOOD BANK SECTION HEAD Chief Medical Technologist Head of the Laboratory OIC-Chief of Hospital
Date: July 16, 2024 Date: July 16, 2024 Date: July 16, 2024 Date: July 16, 2024