LABORATORY STANDARD OPERATING PROCEDURES

FACILITY NAME…………………………………………………

COUNTY…………………………………………………………

SUB COUNTY………………………………………………….

SOP Title: SYPHILLIS TEST SOP No: 025

Version: Original
Effective Date: January 2016 Page 1 of 7

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

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 1. PURPOSE/APPLICABILITY

1.1 PURPOSE: To provide guidelines for the proper procedure of testing for syphilis
antibodies so as to ensure accurate and reliable results.

Applicability: TMH Lab Technologist/Technicians and students

Principle: During the testing procedure, the specimen, serum or plasma is mixed with the
carbon antigen reagent and allowed to react for eight minutes. If anti lipoidal antibodies are
present in the specimen, they will react with the reagent forming visible black floccules. If anti
lipoidal antibodies are not present in the specimen there will be no flocculation.

2. Abbreviations and Terms

2.1 TMH: Tabaka Mission Hospital

2.2 LAB: Laboratory
2.3 TECH: Technologist/Technician
2.4 VDRL: Venereal Disease Research Laboratory

3. Equipment and Materials

3.1 Equipment
3.1.1 Centrifuge
3.1.2 Shaker/mechanical rotor
3.1.3 Slide/Tile
3.1.4 Applicator sticks/stirrer

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 3.1.5 Rubber teats
3.1.6 Needle dropper
3.1.7 Pipettes
3.1.8 Test tubes

3.2 Reagents

3.2.1. Carbon antigen suspension

3.2.2. Positive control
3.2.3. Negative control
3.2.4. Isotonic saline

Reagent storage and stability - Store the reagents at 2-8◦c. Do not freeze. The
shelf life of the reagent is as per the expiry date mentioned on the reagent vial
label. Avoid exposure to elevated temperatures and air as the reagent is highly
sensitive to denaturation and drying.

4. Responsibilities

4.1 The Lab manager is responsible for ensuring the implementation of this SOP with
the assistance of quality officer to ensure strict adherence to this procedure.
4.2 All personnel involved in the execution of this SOP must ensure that they are
properly trained.
4.3 Quality Assurance: SOPs are to be reviewed/approved by the in charge, office of
quality or designee, prior to the responsible approving authority.

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5. Procedures

5.1 Bring all reagents and samples to be tested to room temperature before testing.
Thoroughly mix the carbon antigen reagent suspension by gentle agitation before
testing.
5.2 Qualitative method
1) Place one drop of the test sample positive and negative controls onto
separate reaction circles of the slide using sample dispensing pipette.
2) Add one drop of well-mixed carbon antigen reagent next to the test
sample, positive control and negative control by using a needle dropper
which dispenses 6-18ul of the reagent. Do not let the dropper tip touch
the liquid on the slide.
3) Using a mixing stick, mix the test sample and carbon antigen reagent
thoroughly spreading uniformly over the entire reaction circle.
4) Immediately rotate the slide gently and continuously either manually of on
a mechanical rotor at 1000 r.p.m.
5) Observe fir flocculation macroscopically at 8 minutes.

5.3. Quantitative Method

1) Using isotonic saline prepare serial dilutions of the test sample 1:2, 1:4,
1:8, 1:16, 1:32, 1:62. 1:128 and so on.
2) Perform the qualitative test procedure using each dilution as test
specimen.
3) The titre is reported as the reciprocal of the highest dilution, which shows
a positive test result.

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 5.4 INTERPRETATION OF TEST RESULTS

5.4.1 Qualitative method

a) Large and medium black floccules against white background- Reactive

b) Small black floccules against white background – weekly reactive
c) No floccules - non reactive

• Flocculation is a positive test result and indicates the presence of

anti lipoidal antibodies in the test samples.
• No flocculation is a negative test result and indicates absence of
anti lipoidal antibodies in the test sample.

5.4.2 Quantitative Method- The titre of anti lipoidal antibodies is the highest
dilution of test sample giving a positive test result.

NOTES

1) Samples not tested immediately may be stored at 2-8◦c for up to 48 hours.

2) The reagent and sample dispensing device should dispense precisely 16-18µl and 50µl
respectively.
3) It is strongly recommended that results of the test should be correlated with clinical
findings to arrive at the final diagnosis.
4) Dispose all used and contaminated material as per standard Biohazard safety guidelines.
5) Very slight roughness should be interpreted as a negative test results.
6) Carbon antigen reagent should be gently but thoroughly mixed before testing to dispose
the carbon particles uniformly and improve test readability.

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 7) The reagent contains sodium oxide 0.1% as preservative. Avoid contact with skin and
mucus. On disposal flush with large quantities of water.

6. REFERENCES

AUTHOR TITLE
M.C. Grew B.E. American Journal of Clinical Pathology
Pang Boarn, Mary C. Isolation and purification of serological
active phospholid from breef heart
Monica Cheesbrough Medical Laboratory manual for tropical
countries
Eurocarb Insert

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7. Forms and Appendices

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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