.

LIBEN PRIMARY HOSPITAL

LABORATORY STANDARD
OPERATING PROCEDURE
FOR
H.PYLORI

Compiler: Mekuant&Getachew Signature:

Approver: Abere abye Signature:

Effective Date: Sep. 2013 Revision No. 0

Document No:LPHL/SOP5.5-004 Copy No.

I. REVISION AND AMENDMENT

A. Annual Review of Document
Reviewed by: Approved by:
Revision No. Review Date
Name Signature Name Signature
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B. Version Change History/Description

Version Effectiv Description of Version Change Name & Name &

. No. e Date Signature of Signature of
Reviewer approval

C. Amendment
Rev. Page Description of Amendment Amend Effective Name &
No. No ment Date Signature of
Date approval

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Purpose For proper preparation, detection, identification of H.pylori. Helicobacter

pylori (H. pylori), previously named Campylobacter pyloridis, is a Gram-
negative, microaerophilic bacterium found in the stomach.

Abbreviations
H. pylori = helicobacter pylori
IgM : Immuno globulin M.
IgG : Immuno globulin G.

Material Reagents
1.device
2.desiccant pouch
3.sample dropper

Supplies

1. Cotton swab
2. Disposable gloves
3. Tournicat
4. syringe and niddle
5. vacutaner test tube

NOTE: This is a CONTROLLED document for internal use only.

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6. centrifuge
7. nunck tube
8.Clock,watch,or timing device

9. micro pipette

Sample Sample type Amount required Transport and Stability

Storage
Human serum 0.5ml-1ml 4-30 celicous In case of delay in
testing, sample may
be stored at 2-8 for
maximum up to 24
hours

Safety 1. This package insert must be read completely before performing the test. Failure to
Precautions follow the insert gives inaccurate test results.
2. Do not open the sealed pouch, unless ready to conduct the assay.
3. Do not use expired devices.
4. Bring all reagents to room temperature (15°C-30°C) before use.
5. Do not use the components in any other type of test kit as a substitute for the
components in this kit.
6. Do not use hemolized blood specimen for testing.
7. Wear protective clothing and disposable gloves while handling the kit reagents and
clinical specimens. Wash hands thoroughly after performing the test.
8. Users of this test should follow the US CDC Universal Precautions for prevention
of transmission of HIV, HBV and other blood-borne pathogens.

NOTE: This is a CONTROLLED document for internal use only.

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9. Do not smoke, drink, or eat in areas where specimens or kit reagents are being
handled.
10. Dispose of all specimens and materials used to perform the test as bio hazardous
waste.
11. Handle the Negative and Positive Control in the same manner as patient
specimens.
12. The testing results should be read within 15 minutes after a specimen is applied to
the sample well or sample pad of the device. Read result after 15 minutes may
give erroneous results.
13. Do not perform the test in a room with strong air flow, ie. an electric fan or strong
air-conditioning.
.

Quality Using individual OnSite H. Pylori Ab Rapid Test strip as described in the Assay
Control Procedure below, run 1 Positive Control and 1 Negative Control (provided upon
request) under the
Negative Control
Only the C band shows color development. The T band shows no color development.
Positive Control
Both C and T bands show color development. The appearance of any burgundy color
in the T band, regardless of intensity, must be considered as presence of the band.

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Procedure Step Action

1 Bring the specimen and test components to room temperature if refrigerated
or frozen. Mix the specimen well prior to assay once thawed
2 When ready to test, open the pouch at the notch and remove the test strip.
Place the strip on a clean, flat surface.
3 Fill the plastic dropper with the specimen. Holding the dropper vertically,
dispense 1 drop (about 30-45 μL) of specimen into the sample pad making
sure that there are no air bubbles. Then add 1 drop (about 35 – 50 μL) of
Sample Diluent immediately and wait for 15 minutes
4 Set
5p Results can be read in 15 minutes.
timer

Don’t read result after 15 minutes. To avoid confusion, discard the test device after
result is reported.

Result
Interpretation
1. NEGATIVE RESULT: If only the C band is developed, the test indicates that no
detectable antibodies to H. Pylori are present in the specimen. The result is negative.
2. POSITIVE RESULT: If both C and T bands are developed, the test indicates for
the presence of antibodies to H. Pylori in the specimen. The result is positive.
Samples with positive results should be confirmed with alternative testing method(s)
and clinical findings before a positive determination is made.

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3. INVALID: If no C band is developed, the assay is invalid regardless of color

development on the T band as indicated below. Repeat the assay with a new device.
error and/or that the test reagent has deteriorated. The test should be repeated
using a new strip.

Principle
When an adequate volume of test specimen is applied into the sample pad of the
strip, the specimen migrates by capillary action across the strip. The antibodies:
either the IgG, the IgM, or the IgA, to H. Pylori if present in the specimen will bind
to the H. Pylori conjugates. The immunocomplex is then captured on the membrane
by the pre-coated H. Pylori antigens, forming a burgundy colored T band, indicating
a H. Pylori Ab positive test result. Absence of the T band suggests a negative result.
The test contains an internal control (C band) which should exhibit a burgundy
colored band of the immunocomplex of goat anti-rabbit IgG/rabbit IgG-gold
conjugate regardless the presence of any antibodies to H. Pylori. Otherwise, the test
result is invalid and the specimen must be retested with another device.

Clinical Utility
1. Gastrointestinal diseases included non-ulcer dyspepsia, duodenal and gastric ulcer
and active, chronic gastritis.
2. The prevalence of H.pylori infection could exceed 90% in patients with signs and
symptoms of gastrointestinal diseases. Recent studies indicate an association of H.

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Pylori infection with stomach cancer.

3.H. Pylori colonizing in the gastrointestinal system elicits specific antibody
responses4,5,6 which aids in the diagnosis of H. Pylori infection and in monitoring
the prognosis of the treatment of
4. H. Pylori related diseases. Antibiotics in combination with bismuth compounds
have been shown to be effective in treating active H. Pylori infection
5. H. pylori is associated with clinical improvement in patients with gastrointestinal
diseases providing a further evidence.
6. The OnSite H. Pylori Ab Rapid Test is a latest generation of chromatographic
immunoassay which utilizes recombinant antigens to detect the antibodies to H.
Pylori in human serum or plasma.

Reference
1. Marshall, B.J.et.al.1985. Med. J. Australia. 149:439-44,
2. Soll,A.H. 1990. New England J. Med.322:909-916.
3. Parsonnet, J.et.al.1991. New England J. Med. 325:1127-31.
4. Ansong,R. et.al.1991. J.Clin.Micro. 29:51-53,
5. Pronovost,A.P.et.al. 1994. J.Clin.Microbiol.32:46-50.
6. Megraud, F.et.al.1989. 27:1870-3,1989
7. Marshall,B.J.et.al. 1988. Lancet. Dec.1437-42

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Declaration
.

WE, the undersigned laboratory personnel, certify that we conducting every steps of the procedures
incorporated in this SOP after a prior reading.

Name Signature and Date

1. Abere Abye -----------------------------------------

2. Getachew Abebe -----------------------------------------

3. Mekuant Alemkere -----------------------------------------

4. Girma Zerefaw -----------------------------------------

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