H. pylori Antigen Rapid Test Cassette

(Feces)

Package Insert

REF IHP-602

English

A rapid test for the qualitative detection of Helicobacter pylori (H. pylori) antigens in human feces.

For professional in vitro diagnostic use only.

INTENDED USE

The H. pylori Antigen Rapid Test Cassette (Feces) is a rapid chromatographic immunoassay for the qualitative detection of H. pylori antigens in human feces specimens. It aids in the diagnosis of H. pylori infection.

SUMMARY

H. pylori is a small, spiral-shaped bacterium that lives on the surface of the stomach and duodenum. It is implicated in the etiology of a variety of gastrointestinal diseases, including duodenal and gastric ulcer, non-ulcer dyspepsia, and active and chronic gastritis.

Both invasive and non-invasive methods are used to diagnose H. pylori infection in patients with symptoms of gastrointestinal disease. Specimen-dependent and costly invasive diagnostic methods include gastric or duodenal biopsy followed by urease testing (presumptive), culture, and/or histologic staining. A very common approach to the diagnosis of H. pylori infection is the serological identification of specific antibodies in infected patients. The main limitation of serology tests is the inability to distinguish current and past infections. Antibody may be present in the patient's serum long after eradication of the organism. HpSA (H. pylori Stool Antigen) testing is gaining popularity for diagnosis of H. pylori infection and also for monitoring the efficacy of the treatment of H. pylori infection. Studies have found that more than 90% of patients with duodenal ulcer and 80% of patients with gastric ulcer are infected with H. pylori.

The H. pylori Antigen Rapid Test Cassette (Feces) is a rapid chromatographic immunoassay for the qualitative detection of H. pylori antigens in human feces specimens, providing results in 10 minutes. The test utilizes antibodies specific for H. pylori antigens to selectively detect H. pylori antigens in human feces specimens.

REAGENTS

The test cassette contains monoclonal anti-H. pylori antibodies coated particles and monoclonal anti-H. pylori antibodies coated on the membrane.

PRECAUTIONS

• For professional in vitro diagnostic use only. Do not use after the expiration date.
• The test should remain in the sealed pouch until use.
• Do not eat, drink, or smoke in the area where the specimens or kits are handled.
• Handle all specimens as if they contain infectious agents. Observe established precautions against microbiological hazards throughout all procedures and follow the standard procedures for proper disposal of specimens.
• Wear protective clothing such as laboratory coats, disposable gloves, and eye protection when specimens are assayed.
• The used test should be discarded according to local regulations.
• Humidity and temperature can adversely affect results.

STORAGE AND STABILITY

The kit can be stored at room temperature or refrigerated (2-30°C). The test cassette is stable through the expiration date printed on the sealed pouch. The test cassette must remain in the sealed pouch until use. DO NOT FREEZE. Do not use beyond the expiration date.

SPECIMEN COLLECTION AND PREPARATION

• The feces specimen must be collected in a clean, dry, waterproof container containing no detergents, preservatives, or transport media.
• Bring the necessary reagents to room temperature before use.
• If specimens are to be shipped, they should be packed in compliance with local regulations covering the transportation of etiologic agents.

MATERIALS

Materials Provided:

• Specimen collection tubes with extraction buffer
• Test Cassettes
• Package insert

Materials Required But Not Provided:

• Specimen collection containers
• Pipette and disposable tips (optional)

DIRECTIONS FOR USE

Allow the test cassette, specimen, buffer, and/or controls to reach room temperature (15-30°C) prior to testing.

1. To collect fecal specimens: Collect sufficient quantity of feces (1-2 mL or 1-2 g) in a clean, dry specimen collection container to obtain maximum antigens (if present). Best results will be obtained if the assay is performed within 6 hours after collection. Specimen collected may be stored for 3 days at 2-8°C if not tested within 6 hours. For long term storage, specimens should be kept below -20°C.
2. To process fecal specimens:
   • For Solid Specimens: Unscrew the cap of the specimen collection tube, then randomly stab the specimen collection applicator into the fecal specimen in at least 3 different sites to collect approximately 50 mg of feces (equivalent to 1/4 of a pea). Do not scoop the fecal specimen.
   • For Liquid Specimens: Hold the dropper vertically, aspirate fecal specimens, and then transfer approximately 80 µL into the specimen collection tube containing the extraction buffer. Tighten the cap onto the specimen collection tube, then shake the specimen collection tube vigorously to mix the specimen and the extraction buffer. Leave the tube alone for 2 minutes.
3. Bring the pouch to room temperature before opening it. Remove the test cassette from the foil pouch and use it within one hour. Best results will be obtained if the test is performed immediately after opening the foil pouch.
4. Hold the specimen collection tube upright and open the cap. Invert the specimen collection tube and transfer 2 full drops of the extracted specimen (approximately 80 µL) to the specimen well (S) of the test cassette, then start the timer. Avoid trapping air bubbles in the specimen well (S).
5. Read results at 10 minutes after dispensing the specimen. Do not read results after 20 minutes.

Note: If the specimen does not migrate (presence of particles), centrifuge the extracted specimens contained in the extraction buffer vial. Collect 80 µL of supernatant, dispense into the specimen well (S) of a new test cassette, and start afresh following the instructions mentioned above.

INTERPRETATION OF RESULTS

(Please refer to the illustration above)

POSITIVE: Two colored lines appear. One colored line should be in the control line region (C) and another colored line should be in the test line region (T).

NOTE: The intensity of the color in the test line region (T) will vary depending on the concentration of H. pylori antigen present in the specimen. Therefore, any shade of color in the test line region (T) should be considered positive.

NEGATIVE: One colored line appears in the control line region (C). No line appears in the test line region (T).

INVALID: Control line fails to appear. Insufficient specimen volume or incorrect procedural techniques are the most likely reasons for control line failure. Review the procedure and repeat the test with a new test. If the problem persists, discontinue using the test kit immediately and contact your local distributor.

QUALITY CONTROL

Internal procedural controls are included in the test. A colored line appearing in the control region (C) is an internal valid procedural control. It confirms sufficient specimen volume and correct procedural technique.

Control standards are not supplied with this kit; however, it is recommended that positive and negative controls be tested as a good laboratory practice to confirm the test procedure and to verify proper test performance.

LIMITATIONS

• The H. pylori Antigen Test Cassette (Feces) is for in vitro diagnostic use only. The test should be used for the detection of H. pylori antigens in feces specimens only. Neither the quantitative value nor the rate of increase in H. pylori antigens concentration can be determined by this qualitative test.
• The H. pylori Antigen Test Cassette (Feces) will only indicate the presence of H. pylori in the specimen and should not be used as the sole criteria for H. pylori to be the etiological agent for peptic or duodenal ulcer.
• As with all diagnostic tests, all results must be interpreted together with other clinical information available to the physician.
• If the test result is negative and clinical symptoms persist, additional testing using other clinical methods is recommended. A negative result does not at any time preclude the possibility of H. pylori infection.
• Following certain antibiotic treatments, the concentration of H. pylori antigens may decrease to a concentration below the minimum detection level of the test. Therefore, diagnosis should be made with caution during antibiotic treatment.

EXPECTED VALUES

The H. pylori Antigen Test Cassette (Feces) has been compared with Endoscope-based methods, demonstrating an overall accuracy of 98.6%.

PERFORMANCE CHARACTERISTICS

Sensitivity and Specificity

The H. pylori Antigen Test Cassette (Feces) has been evaluated with specimens obtained from a population of symptomatic and asymptomatic individuals. The result shows that the sensitivity of the H. pylori Antigen Test Cassette (Feces) is 98.8% and the specificity is 98.4% relative to Endoscope-based methods.

Relative Sensitivity: 98.8% (95% CI*: 95.8%-99.9%)

Relative Specificity: 98.4% (95% CI*: 95.5%-99.7%)

Overall Accuracy: 98.6% (95% CI*: 96.8%-99.5%)

Precision

Intra-Assay

Within-run precision has been determined by using 15 replicates of four specimens: negative, low titer positive, middle titer positive, and high titer positive specimens. The specimens were correctly identified >99% of the time.

Inter-Assay

Between-run precision has been determined by 15 independent assays on the same four specimens: negative, low titer positive, middle titer positive, and high titer positive specimens. Three different lots of the H. pylori Antigen Test Cassette (Feces) have been tested using these specimens. The specimens were correctly identified >99% of the time.

Cross-reactivity

Cross reactivity with the following organisms has been studied at 1.0+09 organisms/ml. The following organisms were found negative when tested with the H. pylori Antigen Test Cassette (Feces):

• Acinetobacter calcoaceticus
• Acinetobacter spp
• Branhamella catarrhalis
• Candida albicans
• E. coli
• Chlamydia trachomatis
• Enterococcus faecium
• Enterococcus faecalis
• Gardnerella vaginalis
• Group A Streptococcus
• Hemophilus influenza
• Group B Streptococcus
• Klebsiella pneumonia
• Group C Streptococcus
• Neisseria meningitides
• Proteus mirabilis
• Neisseria gonorrhea
• Proteus vulgaris
• Pseudomonas aeruginosa
• Rotavirus
• Salmonella choleraesius
• Staphylococcus aureus
• Adenovirus

Interfering Substances

The following potentially interfering substances were added to H. pylori negative and positive specimens:

• Ascorbic acid: 20mg/dL
• Oxalic acid: 60mg/dL
• Uric acid: 60mg/dL
• Urea: 2000mg/dL
• Glucose: 2000mg/dL
• Aspirin: 20mg/dL
• Bilirubin: 100mg/dL
• Caffeine: 40mg/dL
• Albumin: 2000mg/dL

BIBLIOGRAPHY

1. Marshall, BJ, McGechie, DB, Rogers, PAR, and Glancy, RG. Pyloric Campylobacter infection and gastroduodenal disease. Med. J. Australia. 1985, 149: 439-444.
2. Soll, AH. Pathogenesis of peptic ulcer and implications for therapy. New England J. Med. 1990, 322: 909-916.
3. Hazell, SL, et al. Campylobacter pyloridis and gastritis I: Detection of urease as a marker of bacterial colonization and gastritis. Amer. J. Gastroenterology. 1987, 82(4): 292-296.
4. Cutler AF. Testing for Helicobacter pylori in clinical practice. Am J. Med. 1996; 100:35S-41S.
5. Anand BS, Raed AK, Malaty HM, et al. Low point prevalence of peptic ulcer in normal individuals with Helicobacter pylori infection. Am J Gastroenterol. 1996, 91:1112-1115.

Vitamin D Rapid Test Cassette

(Whole Blood)

Package Insert

REF OVD-402

English

A rapid test for the semi-quantitative detection of 25-hydroxyvitamin D in human fingerstick Whole Blood.

For professional in vitro diagnostic use only.

INTENDED USE

The Vitamin D Rapid Test Cassette is a rapid chromatographic immunoassay for the semi-quantitative detection of 25-hydroxyvitamin D (25 (OH) D) in human fingerstick Whole blood at a cut-off concentration of 30 ng/mL. This assay provides a preliminary diagnostic test result and can be used to screening for Vitamin D deficiency.

Vitamin D refers to a group of fat-soluble secosteroids responsible for increasing intestinal absorption of calcium, iron, magnesium, phosphate and zinc. In humans, the most important compounds in this group are vitamin D3 and vitamin D2. Vitamin D3 is naturally produced in the human skin through the exposure to ultraviolet light and Vitamin D2 is mainly obtained from foods. Vitamin D is transported to the liver where it is metabolized to 25-hydroxy Vitamin D. In medicine, a 25-hydroxy Vitamin D blood test is used to determine Vitamin D concentration in the body. The blood concentration of 25-hydroxy Vitamin D (including D2 and D3) is considered the best indicator of Vitamin D status. Vitamin D deficiency is now recognized as a global epidemic. Virtually every cell in our body has Receptors for Vitamin D, meaning that they all require sufficient levels of Vitamin D for adequate functioning. The health risks associated with Vitamin D deficiency are far more severe than previously thought. Vitamin deficiency has been linked to various serious diseases: Osteoporosis, Osteomalacia, Multiple Sclerosis, Cardiovascular Diseases, Pregnancy Complications, Diabetes, Depression, Strokes, and higher risks of other serious conditions. Therefore, detecting (25-OH) Vitamin D level is considered a medically necessary screening test, and maintaining sufficient levels not only improves bone health but also enhances overall health and well-being.

The Vitamin D test is an immunoassay based on the principle of competitive binding. During testing, the mixture migrates upward on the membrane chromatographically by capillary action. The membrane is pre-coated with 25 (OH) D antigens on the test line region of the strip. During testing, 25 (OH) D present in the specimen will compete with 25 (OH) D on the test line for a limited amount of anti-25 OH Vitamin D antibodies in the conjugate. The higher the concentration of 25 (OH) D in the specimen, the lighter the T line will be. The result will be read according to the color card provided with the kit. To serve as a procedural control, a colored line will always appear in the control line region indicating that a proper volume of the specimen has been added and membrane wicking has occurred. The test contains anti-25 OH Vitamin D antibody-coated particles and 25-OH Vitamin D antigen coated on the membrane.

PRECAUTIONS

• Please read all the information in this package insert before performing the test.
• For professional in vitro diagnostic use only. Do not use after the expiration date.
• The test should remain in the sealed pouch until ready to use.
• All specimens should be considered potentially hazardous and handled in the same manner as an infectious agent.
• The used test should be discarded according to local regulations.

STORAGE AND STABILITY

Store as packaged at room temperature or refrigerated (2-30°C). The test is stable through the expiration date printed on the sealed pouch or label of the box. The test must remain in the sealed pouch until use. DO NOT FREEZE. Do not use beyond the expiration date.

SPECIMEN COLLECTION AND PREPARATION

The Vitamin D Rapid Test Cassette can be performed using whole blood (from fingerstick).

1. Wash the patient's hand with soap and warm water or clean with an alcohol swab.
2. Massage the hand without touching the puncture site by rubbing down the hand towards the fingertip of the middle or ring finger.
3. Gently prick the finger with a sterile lancet to collect a rounded drop of blood.
4. Add the Fingerstick Whole Blood specimen to the test by using a capillary dropper.
5. Touch the end of the capillary dropper to the blood; do not squeeze the bulb of the dropper. The blood migrates into the dropper through capillary action to the line indicated on the dropper. Avoid air bubbles.
6. Squeeze the bulb to dispense the whole blood to the specimen area of the test cassette. Testing should be performed immediately after the fingerstick Whole Blood has been collected.

MATERIALS

Materials provided:

• Test Cassettes
• Color Card
• Capillary Droppers
• Package Insert

Materials required but not provided:

• Timer
• Buffer
• Lancets

DIRECTIONS FOR USE

1. Allow the test, specimen, buffer and/or controls to reach room temperature (15-30°C) prior to testing.
2. Remove the test cassette from the sealed pouch and use it as soon as possible.
3. Place the cassette on a clean and level surface.
4. To use a capillary dropper: Fill the capillary tube and transfer approximately 20µL of fingerstick whole blood specimen to the specimen area of the test cassette, then add 2 drops of buffer and start the timer. See the illustration below.
5. Wait for the colored line(s) to appear. Read results at 10 minutes by comparing the intensity of the T line with the provided color card. Do not interpret the result after 20 minutes.

Note: It is suggested not to use the buffer beyond 6 months after opening the vial.

INTERPRETATION OF RESULTS

Please refer to the illustration and compare the intensity of the T line with the "Vitamin D Color card" provided with the kit. Do not mix and use Vitamin D color cards from different batches.

25-OH Vitamin D Level	Reference Range (ng/mL)	Reference Range (nmol/L)
Deficient	0-10	0-25
Insufficient	10-30	25-75
Sufficient	30-100	75-250

Deficient: Two colored lines appear. One is in the control region (C) and another should be in the test region (T). The intensity of the line in the test region (T) is equal to the Deficient line (0-10 ng/mL) on the color card, indicating that the Vitamin D level is deficient.

Insufficient: Two colored lines appear. One is in the control region (C) and another should be in the test region (T). The intensity of the line in the test region (T) is equal to the Insufficient line (10-30 ng/mL) on the color card, indicating that the Vitamin D level is insufficient.

Sufficient: Two colored lines appear; one line should always be in the control region (C) and a faint colored line appears in the test region (T). The intensity of the line in the test region (T) is equal to the Sufficient line (30-100 ng/mL) on the color card, indicating that the Vitamin D level is sufficient.

Note: Always compare the intensity of the T line with the "Vitamin D Color card" and interpret results accordingly.

Excess: One colored line appears in the control line region (C). No colored line appears in the test line region (T). This indicates that the Vitamin D level may be excessive, and it is recommended to consult a physician.

Invalid: The control line fails to appear. Incorrect procedural techniques are the most likely reasons for control line failure. Review the procedure and repeat the test with a new test. If the problem persists, discontinue using the test kit immediately and contact your local distributor.

QUALITY CONTROL

An internal procedural control is included in the test. A colored line appearing in the control line region (C) is an internal positive procedural control. It confirms sufficient specimen volume, adequate membrane wicking, and correct procedural technique. Control standards are not supplied with this kit; however, it is recommended that standard controls be tested as a good laboratory practice to confirm the test procedure and verify proper test performance.

LIMITATIONS

• The Vitamin D Rapid Test Cassette provides only a semi-quantitative analytical result. A secondary analytical method must be used to obtain a confirmed result.
• It is possible that technical or procedural errors, as well as other interfering substances in the whole blood specimen, may cause erroneous results.
• The cut-off for the test is 30 ng/mL.
• As with all diagnostic tests, all results must be considered with other clinical information available to the physician.
• Other clinically available tests are required if questionable results are obtained.

EXPECTED VALUES

The Vitamin D Rapid Test Cassette (Whole Blood) has been compared with a predicate device (Vitamin D Rapid Test), demonstrating an overall accuracy of 93.8%.

PERFORMANCE CHARACTERISTICS

Accuracy

The Vitamin D Rapid Test Cassette has been compared with a predicate device (Vitamin D Rapid Test). The following results were tabulated:

Precision

Method Vitamin D Rapid Test Cassette	Predicate Device	(Vitamin D Rapid Test)
Results	Deficient	Insufficient	Sufficient	Total Results
Deficient	4	4	0	8
Insufficient	0	64	2	66
Sufficient	0	0	23	23
Total Results	4	68	25	97
Accuracy	>99.9%	94.1%	92.0%	93.8%

Intra-Assay

Within-run precision has been determined by using 3 replicates of four specimens: 10 ng/mL, 30 ng/mL, 45 ng/mL and 100 ng/mL specimens. The specimens were correctly identified >99% of the time.

Inter-Assay

Between-run precision has been determined by 3 independent assays on the same 4 specimens: 10 ng/mL vitamin D, 30 ng/mL vitamin D, 45 ng/mL vitamin D, 100 ng/mL vitamin D standard samples. Three different lots of the Vitamin D Rapid Test Cassette have been tested using these specimens. The specimens were correctly identified >99% of the time.

Sensitivity and Cross-Reactivity

The Vitamin D Rapid Test Cassette can detect levels of Vitamin D in human fingerstick whole blood as low as 30 ng/mL. The addition of Vitamin A, B, C, E, K and M showed no cross-reactivity.

BIBLIOGRAPHY

1. Holick MF (March 2006). High prevalence of vitamin D inadequacy and implications for health. Mayo Clinic Proceedings. 81 (3): 353-73.
2. Eriksen EF, Glerup H (2002). Vitamin D deficiency and aging: implications for general health and osteoporosis. Biogerontology. 3 (1-2): 73-7.
3. Grant WB, Holick MF (June 2005). Benefits and requirements of vitamin D for optimal health: a review. Alternative Medicine Review. 10 (2): 94-111.
4. Moyad MA. Vitamin D: a rapid review. Dermatol Nurs. 2009, 21:25-30.