LABORATORY STANDARD OPERATING PROCEDURES
FACILITY NAME…………………………………………………
COUNTY…………………………………………………………
SUB COUNTY………………………………………………….
SOP Title: SICKLING TEST SOP No: 024
Version: Original
Effective Date: January 2016 Page 1 of 5
Signatures and Dates:
Author: _ __________________________________________________________________
Laboratory Technologist Date
QA Review: ________________________________________________________________
QA Officer Date
Approving Authority: ________________________________________________________
County Medical Laboratory Coordinator Date
Review/Approval for unchanged documents
DATE Author QA Review Approving Authority
Centre for Health Solutions – Kenya (CHS)
1.0 Purpose & Scope
1.1Purpose
To outline the procedure to be followed when performing the sickling test.
1.2 Scope
This procedure is applicable to Tabaka Mission Hospital Laboratory Haematology section.
1.3 Principle & Safety
1.3.1 Principle
Sodium metabisulfate is a reducing agent and it draws oxygen from the red blood cells
depriving the blood of the oxygen. In the deoxy form, hemoglobin S starts to polymerize
and precipitate out in solution. This distorts the RBC making it rigid and produces the
sickle shaped cell.
1.3.2 Safety;
Always wear personal protective equipment when handling potentially infectious material.
2.0 Abbreviation, Definitions and Terms
2.1 Abbreviations
2.1.1 SOP: Standard Operating Procedure
2.1.2 PMHL: Princess MARINA HOSPITAL LABORATORY
2.2Definitions and Terms
N/A
3.0 Equipment and Materials/Reagents/Supplies
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3.1 Equipment
3.1.1 Microscope
3.2 Materials/Reagents/Supplies
3.2.1 Slides
3.2.2 Cover slip
3.2.3 Paraffin wax
3.2.4 Sodium metabisulfate
3.2.5 Pipette
3.2.6 Pencil
3.2.7 4ml EDTA blood sample
3.2.8 Petroleum jelly
3.2.9 Test tube
4.0 Responsibility
4.1 The Head of section is responsible for monitoring the effective implementation of this
procedure.
4.2 Technical staff is responsible for the implementation of this procedure.
5.0 Procedure
5.1 Procedural steps
5.1.1 Label the slide with the patient number and one of the patient names
5.1.2 Put 2 drops of fresh 2% sodium metabisulfate in the test tube and mix with 1 drop of
well mixed EDTA patient blood.
5.1.3 Gently mix and place a drop onto the slide
5.1.4 Apply a cover slip avoiding creating air bubbles
5.1.5 Create an air tight seal around the cover slip with petroleum jelly
5.1.5.1 Put a small amount of petroleum jelly onto a clean slide
5.1.5.2 Light a burner
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5.1.5.4 Melt the petroleum jelly and quickly drip it onto the edges of the cover slip
creating the airtight seal.
5.1.5.5 Repeat the process until all the four corners of the cover slip are sealed
5.1.6 Check the sealed slide under the microscope. Cells should be in a single layer
5.1.7 Allow to stand for 15-30minuts at room temperature in a wet chamber
5.1.8 Examine the slide under x10 the x40 objectives for sickle cells
5.1.9 Incubate the slide in a wet chamber at 37OC for 6- 24hours
5.1.10 Examine again under the microscope X10 or X 40
5.1.11 Report the results and record in the Peripheral blood smears report book
5.2 Quality control
5.2.1 Use EDTA blood sample
5.2.2 Avoid air bubbles when putting the cover slip
5.2.3 Use freshly prepared 2% sodium metabisulfate
5.2.4 Use a known sickle cell slide as a quality control
5.3 Reporting of results
5.3.1 Report as positive where sickle cells are seen under the microscope x40 objective
5.3.2 Report as negative where red blood cells appear normal at x40 objective
6.0 Related procedures
There are no related procedures
7.0 References
7.1 Heilmeyer L. and Begemann H. Atlas of Clinical Haematology 6th Edition 2005 Library
of Congress Cataloging-in-publication Data.
7.2 Dacie and Lewis Practical Heamatology, 2006 10th Edition, Churchill Livingstone.
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Training log
All members of staff who use this document must date and sign in the table below to indicate
that they have read, understood and are going to implement the document in their work.
Print Name Date Signature
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