Rapid-VIDITEST FOB Blister One Step Fecal Occult Blood Blister test. Instruction manual Producer: VIDIA spol. s r.o., Nad Safinou II 365, Vestec, 252 42 Jesenice, Czech Republic, Tel.: +420 261 090 565, www.vidia.cz INTENDED USE: The Rapid-VIDITEST FOB Blister is a rapid chromatographic immunoassay (non-invasive assay) for the qualitative detection of human hemoglobin in human faeces specimens. To screen for gastrointestinal bleeding, which may be an indicator of colorectal cancer, gastric cancer or peptic ulcers. INTRODUCTION: Colorectal cancer is cancer that occurs in the colon or rectum, and affects both men and women of all racial and ethnic groups, and is most often found in people aged 50 years or older. For men, colorectal cancer is the third most common cancer after prostate and lung cancers. For women, colorectal cancer is the third most common cancer after breast and lung cancers. Blood in the stool may be the only symptom of colorectal cancer, but not all blood in the stool is caused by cancer. Other conditions that can cause blood in the stool include: Hemorrhoids, Anal fissures, Colon polyps, Peptic ulcers, Ulcerative colitis. Gastroesophageal reflux disease (GERD). Crohn's disease, Use of aspirin or nonsteroidal anti-inflammatory drugs (NSAIDs). PRINCIPLE: The Rapid-VIDITEST FOB Blister is a qualitative lateral flow immunoassay for the detection of human hemoglobin in human feces samples. The membrane is pre-coated with monoclonal antibodies against human hemoglobin on the test line region. During testing, the sample reacts with the particle coated with anti-human hemoglobin antibodies which was pre-dried on the test strip. The mixture moves upward on the membrane by capillary action. In the case of a positive result the specific antibodies present on the membrane will react with the mixture conjugate and generate a coloured line. A green coloured band always appears in the control line and serves as verification that sufficient volume was added, that proper flow was obtained and as an internal control for the reagents. MATERALS PROVIDED: - Rapid-VIDITEST FOB Blister tests - Instructions for use - Stool collection tubes-sample diluent 1
MATERIALS REQUIRED BUT NO PROVIDED: - Specimen collection container - Testing tubes or vials - Disposable gloves - Timer SPECIMEN COLLECTION AND PREPARATION: Collect sufficient quantity of faeces (1-2 g or ml for liquid sample). Stool samples should be collected in clean and dry containers (no preservatives or transport media). The samples can be stored in the refrigerator (2-4ºC/36-40ºF) for 1-2 days prior to testing. For longer storage the specimen must be kept frozen at 20ºC/4ºF. In this case, the sample will be totally thawed, and brought to room temperature before testing. PROCEDURES: Specimen preparation: Use a separate specimen collection vial for each sample. 1. Unscrew the cap of the vial and introduce the stick two times into the faecal specimen to pick up quite a lot of sample (50-100mg) (1). Close the vial with the buffer and stool sample. 2. Shake the vial in order to assure good sample dispersion (2). For liquid stool samples, aspirate the faecal specimen with a dropper and add 50-100μL into the specimen collection vial with buffer. TEST PROCEDURE Allow the tests, stool samples and buffer to reach to room temperature (15-30ºC/59-86ºF) prior to testing. Do not open the pack until ready to perform the assay. There are two possibilities for performing the test: A) Using the blister test single pack as a Card test: 1. Cut the blister to obtain a test single pack, hold the non sealed side and open it peeling off the upper foil. Don t remove the test from the blister cavity and use it as soon as possible. 2. Shake the specimen collection tube to assure good sample dispersion. Cut the end of the top (3). 3. Place blister test single pack horizontally and identity it. Dispense exactly 5 drops of Hample+buffer on the white end of the test (4). Read the result at 5 minutes after dispensing the sample. 2
B) Using the blister test single pack as a strip test: By immersion: 1. Cut the blister to obtain a test single pack, hold the non sealed side and open it peeling off the upper foil. 2. Shake the specimen collection vial to assure good sample dispersion. Cut the end of the top (3). 3. Dispense 5 drops of sample+buffer in an identified vial and leave the test strip to stand vertically in the vial (5), taking care of not surpassing the limit of immersion indicated with the arrows. Start the timer. Read the result at 5 minutes. INTERPRETATION OF RESULTS: POSITIVE NEGATIVE INVALID NEGATIVE: Only one GREEN band (control line) appears in the white central zone of the test (control region). POSITIVE: Two lines appears across the central window in the result line region (RED test line marked with the letter T) and in the control line region (GREEN control line marked with the letter C). INVALID: A total absence of the control coloured band (GREEN) regardless of the appearance or not of the result line (RED). Insufficient specimen volume, incorrect procedural techniques or deterioration of the reagents are the most likely reasons for control line failure. Review the procedure and repeat the test performance using a new test. If the problem persists, discontinue using the test kit and contact you local distributor. 3
NOTES ON THE INTERPRETATION OF RESULTS: The intensity of the red coloured band in the result line region (T) will vary depending on the concentration of antigens in the specimen. However, neither the quantitative value, nor the rate of increase in antigens can be determined by this qualitative test. QUALITY CONTROL: Internal procedural controls are included in the test. A green line appearing in the control region is an internal kontrol (C). It confirms sufficient specimen volume and correct procedural technique LIMITATIONS: 1. Rapid-VIDITEST FOB Blister will only indicate the presence of human hemoglobin in the specimen (qualitative detection) and should be used for the detection of hemoglobin in faeces specimens only. Neither the quantitative value nor the rate of increase in hemoglobin concentration can be determined by this test. 2. An excess of sample could cause wrong results (brown bands appear). Dilute the sample with the buffer and repeat the test. 3. Some stool samples can decrease the intensity of the control line. 4. Patients should not collect samples during their menstrual period, if they have bleeding hemorrhoids, blood in urine, or if they have strained during bowel movement. 5. Positive results confirm the presence of hemoglobin in fecal samples; nevertheless, it can be due to several causes, besides colorectal bleeding, such as hemorrhoids, anal fissures, colon polyps, peptic ulcers, ulcerative colitis. gastroesophageal reflux disease (GERD), Crohn's disease. A positive results should be followed up with additional diagnostic procedures by a physician to determine the exact cause and source of the blood in the stool. 6. Negative results do not exclude bleeding since some polyps and colorectal cancers may bleed intermittently or not at all during certain stages of the disease. Additionally, blood may not be uniformly distributed in stool samples. 7. This test may be less sensitive for detecting upper gastrointestinal bleeding because blood degrades as it passes through the gastrointestinal track. EXPECTED VALUES: Colorectal cancer affects both men and women of all racial and ethnic groups, and is most often found in people aged 50 years or older. For men, colorectal cancer is the third most common cancer after prostate and lung cancers. For women, colorectal cancer is the third most common cancer after breast and lung cancers. PERFORMANCE CHARACTERISTICS: Sensitivity and specificity The evaluation was conducted comparing the results obtained using the Rapid-VIDITEST FOB Blister to a commercial available guaiac assay. Rapid-VIDITEST FOB Blister was highly specific (>99%) and also highly sensitive (>99%) compared with the results of that guaiac assay. The detection of hhb with Rapid-VIDITEST FOB Blister showed >99% of sensitivity and >99% of specificity compared with others commercial rapid tests (ImmunoTech OcculTech and Human Hexagon OBTI). 4
Cross-reactivity It was performed an evaluation to determine the cross reactivity and interference of Rapid-VIDITEST FOB Blister. There is not cross reactivity with common gastrointestinal pathogens, other organism and substance occasionally prezent in feces: Rotavirus, Astrovirus, Adenovirus, Escherichia coli, Campylobacter, Giardia lamblia. STORAGE AND STABILITY: Store as packaged in the sealed pouch either at refrigerated or room temperature (2-30ºC/36-86ºF). The test is stable through the expiration date printed on the sealed pouch. The test must remain in the sealed pouch until use. Do not freeze. PRECAUTIONS: - For professional in vitro diagnostic use only. - Do not use after expiration date. - The test should remain in the sealed pouch until use. - Do not use the test if pouch is damaged. - Follow Good Laboratory Practices, wear protective clothing, use disposal gloves, do not eat, drink or smoke in the area. - All the specimens should be considered potentially hazardous and handled in the same manner as an infectious agent. - The test should be discarded in a proper biohazard container after testing. - The test must be carried out within 2 hours of opening the sealed bag. REFERENCES: 1. WALKER C.W., Fecal occult blood tests reduce colorectal cancer mortality., Am Fam Physician. 2007 Jun 1;75(11):1652-3. SYMBOLS FOR IVD COMPONENTS AND REAGENTS: In vitro diagnostic device Use by Batch code Manufacturer 5