Standard Operating Procedure SOP # 0506

Antibody Identification

Procedure Custodian Biomedical Science Program

Approval Authority Program Coordinator

Program Coordinator Martin Gonzo Signature:

Status Final

Procedure created by Edwig Hauwanga Date created:

Procedure reviewed by S Blaauw Signature:

Date Reviewed Procedure Approved 25.01.2016

Date Scheduled for next review 2018

Contact Person(s) details S Blaauw Tel no: 061 207 2970

VERSION CONTROL AND CHANGE HISTORY

Version Control Date Approved Approved by PC Amendment

#1 2017 Reviewed

 All safety procedures for specimen handling and disposal must be adhered to
 All microbiological samples for processing should be considered a potential source of
transmissible infections so universal safety precautions should be observed.
 Hands should be thoroughly washed with soap and water before and after handling all
samples.
 Disinfect all work surfaces prior to testing and after processing
 All samples and culture plates should be properly discarded according to the current
standard for disposal of hazardous waste.
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 All patient samples should be treated as potentially infectious and handled according to
standard precautions.
 Wear laboratory coats and gloves when processing and analyzing specimens.
 Never pipette by mouth.
 After working with blood/plasma specimens remove gloves and wash hands with soap and
water.
 Use biocide to disinfect spills. Use biocide to decontaminate pipette tips and glass tubes.

This SOP is mainly for students to familiarise them with the principle and the expected results
and values when this test is performed.

1. PURPOSE

This technique is to be used in identification of the antibodies of specimens with a positive

antibody screen.

2. SCOPE OF APPLICATION

 This procedure should be used by a qualified technologist for the identification of

unexpected antibodies in specimens yielding a positive result for antibody screening
 It can be used to train students and trainees as well

3. DEFINITIONS

Define any jargon or acronyms used in this procedure/guideline document.

VERSION CONTROL AND CHANGE HISTORY

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4. PROCEDURES/GUIDELINES

Specimen

 Clotted sample in a red top tube (please avoid gel top tube specimens)

Regents and equipment

Panel cells
Broad spectrum AHG
LIR

Test tube (12x75ml)

Tube rack
Pipettes

Serological centrifuge

Procedure

1. Number tubes from 1-9 and the patient identification number.

2. Using a Pasteur pipette, transfer 2 drops of patient serum into the each of the tubes.
3. Drop the panels into the tubes according to the numbers
4. Add 2 drops of LIR in each tube and spin @1000rpm for 1 minute and record results in
IS section
5. Incubate tube @37 in the water bath for 10-30 minutes
6. Remove tubes from water bath, spin @1000rpm and record your results in the “37”
section
7. Wash the cells by filling the tubes with ¾ pf saline and then centrifuging the tubes @3000
for 1 minute
8. Decant the saline, resuspend the cells and repeat washing twice more
9. After the 3rd wash decant the saline completely
10. Resuspend the cells and add AHG
11. Centrifuge the tubes @2000 rpm for 30 seconds
12. Remove from centrifuge, read for agglutination and record test results at the “AHG phase
“ section on the worksheet
13. Control the negative tubes by adding sensitized cells, centrifuge @2000 rpm for 30
seconds and read and record results at the “control” section
14. Use the decode sheet to interpret results.

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5. REFERENCES

 NAMBTS SOP: Methods for performing standard laboratory tests

6. APPENDICES

 NB! You will need an antibody decode sheet for specific batch lot screen cells being
used to interpret results.

Worksheet

Panel 1 2 3 4 5 6 7 8 9 AA
IS
37
IAT
Control

Scanned Copy of the decode

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PLEASE NOTE: All students relevant staff must familiarize themselves with this
procedure and have to sign on this page that they understand the content of this
procedure. Students must be able to answer questions on this procedure.

Document: read by Signature Date

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PLEASE NOTE: All students must familiarize themselves with this procedure and have to sign on this page that
they understand the content of this procedure. Students must be able to answer questions on this procedure.

Document read by Signature Date

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