Addis Ababa City Administration Health Bureau
Zewditu Memorial Hospital
Laboratory Case team
Title: KHB Effective date: July 2, 2012
Document type: SOP Next revision date.: July 1, 2013
Document Number: P-017/12 Version No.: 01
Section/Subject: Serology Page: 1 of 7
Purpose For the screening of donated blood And as an aid in assessing the clinical
condition of persons with unknown status related to infection with HIV
HIV………………human immune deficiency virus
Abbreviations AIDS ……………. Acquired immunodeficiency syndrome
ELISA …………… Enzyme linked immunosorbent assay
C…………………. control
T…………………. Test
Ags ………………. Antigens
Abs ……………….. Antibodies
Materials Reagents list
HIV (1+2) antibody cassettes
Sample diluents
Sodium hypochlorite solution (5 %) or other suitable disinfectant.
Reagents, stability and storage: At room temperature
Supplies
Disposable gloves
biohazard waste containers
Timer
Pipette tips and pipettes
Sample Sample type Amount required Transport and Stability
Storage
K3 EDTA blood 4-5ml Room Use fresh blood
N.B:- reject less Temperature
NOTE: This is a CONTROLLED document for internal use only
Addis Ababa City Administration Health Bureau
Zewditu Memorial Hospital
Laboratory Case team
Title: KHB Effective date: July 2, 2012
Document type: SOP Next revision date.: July 1, 2013
Document Number: P-017/12 Version No.: 01
Section/Subject: Serology Page: 2 of 7
than 1/3 ml of the
standard collection
tube.
Serum 2.0 ml 2.-8 Oc One week
>-20 For log period
Plasma 2.0 ml 2.-8 Oc One week
>-20 Oc For log period
Limitation 1. The HIV-1/HIV-2 test is designed to detect antibodies to HIV-1 and HIV-2 in
human serum, plasma and whole blood. Other body fluids or pooled samples
may not give accurate results.
2. No test absolutely guarantees that a sample does no contain low level of
antibodies to HIV-1 and HIV-2 which may occur at a very early stage of
infection. Therefore a negative result should not exclude the possibility of
exposure to or infection by HIV-1 or HIV-2 viruses.
3. Positive samples should be retested, and additional testing using other
clinical methods is recommended
Safety Using universal persecution (gloves, lab coat, washing hands) when handling
Precautions infectious materials refer to the national health and safety guideline for standard
safety procedure
Maintenance Step Action
Daily Cleaning
Quality Control Level Stability Frequency Preparation
Control (y/n)
Internal kit Sero-03 Room Each run N
control temperature
NOTE: This is a CONTROLLED document for internal use only.
Addis Ababa City Administration Health Bureau
Zewditu Memorial Hospital
Laboratory Case team
Title: KHB Effective date: July 2, 2012
Document type: SOP Next revision date.: July 1, 2013
Document Number: P-017/12 Version No.: 01
Section/Subject: Serology Page: 3 of 7
In house Sero-03 -20 or Each run Y
control colder
Step Action
Procedure 1 Remove a test cassette from a foil pouch and place it on a flat
surface
2 Use the sample either serum/plasma or whole blood: Add 40µl of
sample (precision pipette) to the sample area
3 Add one drop (~40µl) of sample diluents to the same area
4 The result can be seen within 2-3 minutes with strong positive
samples. Please do not interpret the test results after 30 minutes. Do
record the result on cassette.
Result 1. Positive result (two bands)
Interpretation A reddish-purple band appears both on control line (C-line) and test line (T-line)
of the cassette.
2. Negative result (one band)
A reddish-purple band appears only at the control line (C-line) of the cassette.
3. Invalid result
No reddish-purple band appears neither at the control line nor the test line of the
cassette
If there is no distinct pink/purple line visible in the CONTROL (C) area
Any lines that appear outside of the Control (C) Area or Test (T) Area
NOTE: This is a CONTROLLED document for internal use only.
Addis Ababa City Administration Health Bureau
Zewditu Memorial Hospital
Laboratory Case team
Title: KHB Effective date: July 2, 2012
Document type: SOP Next revision date.: July 1, 2013
Document Number: P-017/12 Version No.: 01
Section/Subject: Serology Page: 4 of 7
Principle The gold-gp160 conjugate and gold-gp36 conjugate are coated to the conjugate pad in
advance. The test line (HIV type I+II antigens) and the control line (monoclonal antibody
against gp160) and pre-coated on the surface of NC membrane. When the sample that
added to the sample pad migrate through the conjugate pad, it reconstitutes and mixes
with colloidal gold-antigen conjugates. The mixture continues to migrate through the NC
membrane to the pre-coated antigens or antibody present on the membrane. A purple red
test line will be visible in the strip if there are enough antibodies to HIV-1/HIV-2 in the
sample. If antibodies to HIV-1/ HIV-2 are absent, or are present at very level, then no
color will appear in the test line. The control line purple red is used as quality control
only and does not affect the result of the test.
1. Gallo, RC et al. Detection and Isolation of Cytopathic Retroviruses [HTLV-
Reference III] from patients with AIDS and at risk for AIDS. Science 1984; 224:500-
503.
NOTE: This is a CONTROLLED document for internal use only.
Addis Ababa City Administration Health Bureau
Zewditu Memorial Hospital
Laboratory Case team
Title: KHB Effective date: July 2, 2012
Document type: SOP Next revision date.: July 1, 2013
Document Number: P-017/12 Version No.: 01
Section/Subject: Serology Page: 5 of 7
2. Spielberg F, Kabeya CM, Ryder RW, et al. Field testing and comparison
evaluation of rapid, visually read screening assays for antibody to human
immunodefiency virus. Lancet, 1989; 1:580-584.
3. Osikowicz G, Begge M. One step chromatographic immunoassay for
quantitative determination of choriogonadotropin in urine. Clin. Chem, 1990,
36:1586.
4. Brandt J, Andersson LO, Porath attachment of proteins to polysaccharide
carriers by means of benzoquinone. Biochim Biophys Acta, 1975, 386:196-
202.
NOTE: This is a CONTROLLED document for internal use only.
Addis Ababa City Administration Health Bureau
Zewditu Memorial Hospital
Laboratory Case team
Title: KHB Effective date: July 2, 2012
Document type: SOP Next revision date.: July 1, 2013
Document Number: P-017/12 Version No.: 01
Section/Subject: Serology Page: 6 of 7
Name Signature
Prepared by: ------------------------ -----------------------------
Reviewed by: --------------------------- -----------------------------
Approved by: -------------------------- ------------------------------
NOTE: This is a CONTROLLED document for internal use only.
Addis Ababa City Administration Health Bureau
Zewditu Memorial Hospital
Laboratory Case team
Title: KHB Effective date: July 2, 2012
Document type: SOP Next revision date.: July 1, 2013
Document Number: P-017/12 Version No.: 01
Section/Subject: Serology Page: 7 of 7
Declaration
I, the undersigned laboratory personnel, certify that I am conducting every steps of the procedures
incorporated in this SOP after a prior reading.
Name Signature and Date
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NOTE: This is a CONTROLLED document for internal use only.
