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OnSite H. pylori Ag Rapid Test - Cassette (Fecal Specimen) Page 1 of 2

Collection stick

OnSite™

H. pylori Ag Rapid Test

R0192C

INTENDED USE

The OnSite H. pylori Ag Rapid Test is a lateral flow chromatographic immunoassay for the

qualitative detection of H. pylori antigen in human fecal specimen. It is intended to be used by

professionals as a screening test and provides a preliminary test result to aid in the diagnosis

of infection with H. pylori.

Any interpretation or use of this preliminary test result must also rely on other clinical findings

as well as on the professional judgment of health care providers. Alternative test method(s)

should be considered to confirm the test result obtained by this device.

SUMMARY AND EXPLANATION OF THE TEST

Helicobacter pylori (H. pylori) a gram-negative, helical, rod-shaped bacterium, colonizes the

gastric mucosa of approximately one-half of the world population

. H. pylori infection is a risk

factor for a variety of gastrointestinal diseases including non-ulcer dyspepsia, duodenal and

gastric ulcers and active, chronic gastritis

2-6

. Therefore elimination of H. pylori may be the most

promising strategy to reduce the incidence of gastric cancer

H. pylori can be transmitted through the ingestion of food or water that is tainted with fecal

matter. Antibiotics in combination with bismuth compounds have been shown to be effective in

treating active H. pylori infection.

H. pylori infection is currently detected by invasive testing methods based on endoscopy and

biopsy (i.e. histology, culture) or non-invasive testing methods such as Urea Breath Test

(UBT), serologic antibody test and stool antigen test. UBT has a high accuracy but requires

expensive lab equipment and use of a radioactive reagent

. Serologic antibody tests detect

IgG specific to H. pylori, and cannot distinguish between current active infections and past

infections. The stool antigen test detects antigen present in the feces, which indicates an

active H. pylori infection. It can also be used to monitor the effectiveness of treatment and the

recurrence of an infection, and is not affected by the use of Proton Pump Inhibitors (PPI)

The OnSite H. pylori Ag Rapid Test detects H. pylori antigen present in the fecal specimen by

using specific antibodies. The test can be performed within 10 minutes by minimally skilled

personal without the use of laboratory equipment.

TEST PRINCIPLE

The OnSite H. pylori Ag Rapid Test is lateral flow

chromatographic immunoassay. The test strip in

the cassette device consists of: 1) a burgundy

colored conjugate pad containing anti-H.pylori

specific antibody conjugated with colloidal gold

(anti-H. pylori conjugate) and 2) a nitrocellulose

membrane strip containing a test line (T line) and a

control line (C line). The T line is pre-coated with anti-H. pylori antibody, and the C line is pre-

coated with a control line antibody.

When an adequate volume of extracted fecal specimen is dispensed into the sample well of

the cassette, the specimen migrates by capillary action across the cassette. The H. pylori

antigen, if present in the specimen, will bind to the anti-H. pylori conjugate. The

immunocomplex is then captured on the membrane by the pre-coated antibody forming a

burgundy colored T line, indicating an H. pylori Ag Rapid Test positive result.

Absence of the T line suggests an H. pylori Ag Rapid Test negative result. The test contains an

internal control (C line) which should exhibit a burgundy colored line of the immunocomplex of

the control antibodies regardless of the color development on the T line. If no control line (C

line) develops, the test result is invalid and the specimen must be retested with another device.

REAGENTS AND MATERIALS PROVIDED

1. Individually sealed foil pouches containing:

a. One cassette device

b. One desiccant

2. Stool collection devices, each containing 1 mL Sample Extraction Buffer (REF SB-

R0192)

3. Plastic droppers for transferring watery stool

4. Patient ID stickers

5. One package insert (instruction for use)

MATERIALS MAY BE REQUIRED AND AVAILABLE FOR PURCHASE

1. Positivia H. pylori Ag Rapid Test Control Kit (Cat # C0192) contains positive control and

negative control

MATERIALS REQUIRED BUT NOT PROVIDED

1. Clock or timer

2. A container to hold fecal specimen

WARNINGS AND PRECAUTIONS

For in Vitro Diagnostic Use

1. This package insert must be read completely before performing the test. Failure to

follow the insert causes inaccurate test results.

2. Do not open the sealed pouch, unless ready to conduct the assay.

3. Do not use any kit components beyond their stated expiration dates.

4. Do not use the components from any other type of test kit as a substitute for the

components in this kit.

5. Bring all reagents to room temperature (15-30°C) before use.

6. Do not scoop stool sample as this may lead to excess fecal specimen that tends

to clog the sample pad and interfere with sample migration.

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 bio-safety.

9. Do not smoke, drink or eat in areas where specimens or kit reagents are being handled.

10. Avoid extraction buffer contact with skin or eyes. Do not ingest.

11. Dispose of all specimens and materials used to perform the test as bio-hazardous

waste.

12. The test results should be read 10-15 minutes after a specimen is applied to the

sample well of the device. Any results interpreted outside of the 10-15 minute

window should be considered invalid and must be repeated.

13. Do not perform the test in a room with strong air flow, i.e. electric fan or strong air-

conditioning.

REAGENT PREPARATION AND STORAGE INSTRUCTIONS

All reagents are ready to use as supplied. Store unopened test devices at 2-30°C. If stored at

2-8°C, ensure that the test device is brought to room temperature before opening. The test

device is stable through the expiration date printed on the sealed pouch. Do not freeze the kit

or expose the kit to temperatures above 30°C.

SPECIMEN COLLECTION AND HANDLING

Consider any materials of human origin as infectious and handle them using standard bio-

safety procedures.

To prepare specimens using solid fecal samples follow Procedure A below. To prepare

specimens using watery fecal samples follow Procedure B below.

Procedure A: Solid fecal specimens

Step 1: Collect a random stool specimen in a clean, dry receptacle.

Step 2: Label the stool collection device with the specimen’s ID number (patient ID sticker).

Open the stool collection device by unscrewing the top and use the collection stick to

randomly pierce in 2-5 different sites, twisting the collection stick into the fecal

specimens to help collection if necessary. Do not scoop fecal specimen as this

may lead to an invalid test result.

Step 3: Ensure that all inner grooves of the collection stick are filled with fecal specimen.

However, excess fecal specimen on the outside of grooves should be scraped off.

Incorrect sampling may lead to an erroneous test result.

Step 4: Replace the collection stick and tighten securely to close the stool collection device.

Step 5: Shake the stool collection device vigorously.

The specimen is now ready for testing, transportation or storage.

Procedure B: Watery fecal specimens

Step 1: Collect a random fecal specimen in a clean, dry receptacle.

Step 2: Label the stool collection device with the specimen’s ID number (patient ID sticker).

Open the stool collection device by unscrewing the top.

Step 3: Fill the plastic dropper with the specimen; dispense 2 drops (70-85 µL) into the stool

collection device.

Step 4: Replace the collection stick and tighten securely to close the stool collection device.

Step 5: Shake the stool collection device vigorously.

The specimen is now ready for testing, transportation or storage.

Note: The extracted specimens may be stored at 2-8°C or at room temperature up to 37°C

for 10 days. For longer storage, the extracted specimen may be frozen at -20°C.

Avoid multiple freeze-thaw cycles.

TEST PROCEDURE

Step 1: Bring the specimen and test components to room temperature if refrigerated or

frozen. Once the specimen is thawed, mix well prior to performing the assay.

Step 2: When ready to test, open the pouch at the notch and remove the test device. Place

the test device on a clean, flat surface.

Step 3: Shake the stool collection device vigorously to ensure a homogenous liquid

suspension.

Step 4: Hold the stool collection device vertically. Twist off the cap. Dispense 2 drops (70-

90 µL) of the solution into the sample well of the cassette. Do not overload the

solution.

Step 5: Set up the timer.

Step 6: Results can be read at 10 minutes. Positive results can be visible in as short as 1

minute. Negative results must be confirmed at the end of the 15 minutes only.

However, any results interpreted outside of the 10-15 minute window should

be considered invalid and must be repeated. Discard used device after

interpreting the result following local requirements governing the disposal of

device.

Collection stick

2 drops

Dispenser cap

10 minutes

Result

2 drops of solution

2-5 sites

OnSite H. pylori Ag Rapid Test - Cassette (Fecal Specimen) Page 2 of 2

QUALITY CONTROL

1. Internal Control: This test contains a built-in control feature, the C line. The C line

develops after adding specimen extract. If the C line does not develop, review the entire

procedure and repeat the test with a new device.

2. External Control: Good Laboratory Practice recommends using external positive and

negative controls to assure the proper performance of the assay, particularly under the

following circumstances:

a. A new operator uses the kit, prior to performing testing of specimens.

b. A new lot of test kits is used.

c. A new shipment of test kits is used.

d. The temperature during storage falls outside of 2-30°C.

e. The temperature of the test area falls outside of 15-30°C.

f. To verify a higher than expected frequency of positive or negative results.

g. To investigate the cause of repeated invalid results.

INTERPRETATION OF ASSAY RESULT

1. NEGATIVE RESULT: If only the C line develops, the test indicates that no detectable H.

pylori antigen is present in the specimen. The result is negative or non-reactive.

2. POSITIVE RESULT: If both C and T lines develop, the test indicates the presence of

detectable H. pylori antigen in the specimen. The result is positive or reactive.

Fecal specimens with positive results should be interpreted in conjunction with other

testing procedures and clinical findings before a diagnosis is made.

3. INVALID: If no C line develops, the assay is invalid regardless of any color

development on the T line as indicated below. Repeat the assay with a new test device.

Excess fecal specimen can lead to invalid test results; if this is the cause, re-

sample and re-test (see instructions for collection of specimen).

PERFORMANCE CHARACTERISTICS

1. Clinical Performance

A total of 157 fecal specimens were collected from symptomatic patients and healthy

individuals. Specimens were tested by the OnSite H. pylori Ag Rapid Test. The urea

breath test (UBT) gold standard is used as the reference test method. Comparison for

all subjects is shown in the following table:

Relative Sensitivity: 96.7%, Relative Specificity: 93.8%, Overall agreement: 94.9%

2. Analytic Sensitivity

Six groups of fecal specimen extracts from 20 healthy individuals were spiked with H.

pylori lysate antigen (Strain 43504) at concentrations of 0, 0.25, 0.5, 0.75, 1, and

2 ng/mL, respectively, and tested with the OnSite H. Pylori Ag Rapid Test. The results

were showed in the follow table. The detection limit of the OnSite H. pylori Ag Rapid

Test as defined as the level of ≥95% positive detection is 1 ng/mL of H. pylori lysate

antigen.

H. pylori Lysate Antigen (ng/mL)

0 0.25 0.5 0.75 1 2

Number of positive 0 0 0 9 20 20

Number of negative 20 20 20 11 0 0

Detection rate % 0% 0% 0% 45% 100% 100%

n=20, relative sensitivity at 1 ng/mL is 100%

3. Cross-Reactivity

The organisms listed below were tested for cross-reactivity with the OnSite H. Pylori Ag

Rapid Test. No cross-reactivity was observed on the organisms at ≥ 1 x 10

org/mL.

cinetobacter calcoaceticu

Neisseria gonorrheae

denoviru

Neisseria meningitides

Enterococcus faecalis Proteus mirabilis

Escherichia coli Proteus vulgaris Hauser

Gardnerella vaginalis Pseudomonas aeruginosa

Geotrichum candidum Rotavirus

Haemophilus influenza Salmonella Paratyphi A

α-haemolytics streptococcus Salmonella Paratyphi B

Β-haemolytic streptococcus Salmonella Paratyphi C

Klebsiella pneumonia Salmonella typhi

Moraxella catarrhalis

4. Interference

The following common and potentially interfering substances may affect the

performance of the OnSite H. Pylori Ag Rapid Test. This was studied by spiking these

substances into negative and positive fecal specimens, respectively. The results

demonstrate, at the concentrations tested, the substances studied do not affect the

performance of the OnSite H. Pylori Ag Rapid Test.

List of potentially inferring substances and concentrations tested:

ums® Antacid

mg/mL Pepto-Bismol® Antacid 1:20

agamet® Antacid

mg/mL Barium sulfate

Prilosec® Antacid

mg/mL Hemoglobin (tarry stool) 12.5%

Mylanta® Antacid 1:20

LIMITATIONS OF THE TEST

1. The Assay Procedure and the Interpretation of Assay Result sections must be followed

closely when testing for the presence of H. pylori antigen in feces. Failure to follow the

procedure, particularly the Specimen Collection and Handling procedure, may lead to

inaccurate results.

2. The OnSite H. pylori Ag Rapid Test is limited to the qualitative detection of H. pylori

antigen in human fecal specimen. The intensity of the test line does not have a linear

correlation with the antigen titer in the specimen.

3. A negative or non-reactive result indicates the absence of detectable H. pylori antigen.

However, a negative test result does not preclude the possibility of infection with H.

pylori.

4. A negative or non-reactive result can occur if the quantity of the H. pylori antigen

present in the specimen is below the detection limits of the assay or if the antigens that

are detected are not present in the fecal specimen collected.

5. It is reported that the seroprevalence of H. pylori in specimens with positive fecal occult

blood (FOB) test results is approximately 39.3%

. Therefore a specimen that tests

positive with an FOB test may also be tested positive with the OnSite H. pylori Ag Rapid

Test.

6. If symptoms persist and the result from the OnSite H. pylori Ag Rapid Test is negative

or non-reactive, it is recommended to test with alternative test methods.

7. The results obtained with this test should only be interpreted in conjunction with other

diagnostic procedures and clinical findings.

REFERENCES

1. Fashner J, Gitu AC. Diagnosis and Treatment of Peptic Ulcer Disease and H.pylori

infection. Am Fam Physician. 2015 Feb 15;91(4):236-42

2. Asaka M, Kato M, Takahashi S, et al. Guidelines for the management of Helicobacter

pylori infection in Japan: 2009 revised edition. Helicobacter 2010; 15:1–20.

3. Fischbach W, Malfertheiner P, Hoffmann JC, et al. S3-guideline “helicobacter pylori and

gastroduodenal ulcer disease” of the German society for digestive and metabolic

diseases (DGVS) in cooperation with the German society for hygiene and microbiology,

society for pediatric gastroenterology and nutrition e. V., German society for

rheumatology, AWMF-registration-no.021/001. Z Gastroenterol 2009;47:1230–63.

4. Fock KM, Talley N, Moayyedi P, et al. Asia-Pacific consensus guidelines on gastric

cancer prevention. JGastroenterolHepatol 2008;23:351–65.

5. Malfertheiner P, Bornschein J, Selgrad M. Role of Helicobacter pylori infection in gastric

cancer pathogenesis: a chance for prevention. J Dig Dis 2010;11:2–11.

6. Polk DB, Peek RM Jr. Helicobacter pylori: gastric cancer and beyond. Nat Rev

Cancer 2010;10:403–14.

7. Malfertheiner P, Megraud F, O'Morain CA, et al. European Helicobacter Study Group.

Management of Helicobacter pylori infection—the Maastricht IV/ Florence Consensus

Report. Gut 2012;61:646–64.

8. Shimoyama T, Kato T, Kodama M, et al. Applicability of a monoclonal antibody-based

stool antigen test to evaluate the results of Helicobacter pylori eradication therapy. Jpn J

Infect Dis 2009. 62(3):225-7.

9. Peter M, Francis M, Colm AO, et al. Management of Helicobacter pylori infection-the

Maastricht IV/ Florence Consensus Report. Gut. 2012 May;61(5):646-64.

10. Ugwuja E, Ugwu N. An Assessment of Faecal Occult Blood Test and H. pylori infection

in Patients with Uninvestigated Dyspepsia in Primary Health Cares in Abakaliki, Nigeria.

The Internet J of laboratory Medicine 2003 V3 No. 1

OnSite H. Pylori Ag Rapid Test

UBT reference Positive Negative Total

Positive 58 2 60

Negative 6 91 97

Total 64 93 157

CTK Biotech, Inc.

10110 Mesa Rim Road

San Diego, CA 92121,USA

Tel: 858-457-8698

Fax: 858-535-1739

E-mail: [email protected]

For Export Only. Not For Resale In The USA

S S

SIMPLIFYING DIAGNO STICS

Index of CE Symbols

Consult

instructions for use

For in vitro

diagnostic use only

Use by

Catalog #

Lot Number

Tests per kit

Store between 2-30°C

Authorized

Representative

Do not reuse

Manufacturer

Date of manufacture

MDSS GmbH

Schiffgraben 41

30175 Hannover, Germany

PI-R0192C Rev. F

Date released: 2017-09-19

English version