ONE STEP Fecal Occult Blood TEST

(8) Do not mix and interchange different specimen. (9) Care should be taken to avoid contamination of the end of bottle when dropping of assay diluent into sample well. (10) SD BIOLINE FOB assay diluent contain a proprietary anti-microbial agent which presents no hazard to the user if normal laboratory safety precautions are followed.

5. Specimen collection, Preparation, Storage and Precaution 1. Explanation of the test

[Introduction] The detection of fecal occult blood is important for the diagnosis of disease that results in gastrointestinal bleeding and to screen for colorectal cancers and large adenomas that bleed. Screening for colorectal cancer probably increases the cancer detection at an early stage, therefore reduces the mortality. Earlier commercially available FOB chemical test has the inherent problems of false reaction that result from diet prior to sample collection. To overcome these potential problems in chemical test, human hemoglobin specific monoclonal antibodies is used as capture and detector materials in SD BIOLINE FOB test. [Test principle] SD BIOLINE FOB Test is designed to detect human hemoglobin in fecal samples. SD BIOLINE FOB test has a membrane strip, where are two pre-coated lines as T (Test Line), C (Control Line) on the surface of the test device. Two lines in result window are not visible before applying any samples. The Control Line is used for procedural control. Control line should always appear if the test procedure is performed properly and the test reagents of control line are working. A purple T line will be visible in the result window if there is enough hemoglobin in the sample. If hemoglobin is not present in the sample, there is no color appearance in T line. After specimen collection and preparation, a extracted sample is added to the test, and then hemoglobin in extracted sample react with anti-human hemoglobin of colloidal gold conjugates pad and forms a complex of hemoglobin and colloidal gold conjugates. As this mixture migrates along the length of the membrane by capillary action, the hemoglobin and colloidal gold conjugates complex is captured by the relevant another anti-human hemoglobin immobilized in test line across the test device and generate a colored line. [Intended Use] SD BIOLINE FOB kit is a rapid, qualitative test for the detection of human blood hemoglobin in human feces specimens. SD BIOLINE FOB kit is intended for professional use, only for an initial screening test and reactive samples should be confirmed by a supplemental assay such as a commercial clinical chemistry method. This kit is for in vitro diagnostic use only. 1) Sample collection and preparation (1) Collect a random sample of feces in clean, dry container or receptacle, making sure to exclude urine. (2) Loosen Square-shaped cap on the upper part of sample collection tubes and remove applicator stick. Take care not to spill or splatter solution from the tubes and not to loosen cap on the lower part of the sample collection tube. (3) Collect random sample by using the applicator stick in the proper method, that is, insert the applicator stick and turn the stick into the fecal sample several times (5 ~ 6 times) at different sites so as to get a representative sampling. Fill up the ditch with fecal sample and please check its quantity whether too much or not. (4) Re-insert the applicator stick into the sample collection tube and screw the white cap tightly. (5) Shake and mix well the tube vigorously to provide a good homogenous extraction and suspension. 2) Specimen transport and storage / precaution (1) Fresh fecal specimen should be used for this FOB test. (2) Fecal specimen should be tested as soon as possible after collection. Do not use any kind of transport media to store or transport the specimens. (3) Fecal sample to be colleted into container should be tested within 72 hours at room temperature. (4) Fecal sample to be extracted are stable for up to 3 days at room temperature. For storage periods greater than 3 days, refrigeration is recommended. (5) If the test is not performed with this extracted fecal sample immediately, the collected and prepared fecal sample into collection tube is now ready to be stored at 2 ~8 to transport or test.

Invalid Result : No line appears. If the purple color band is not visible within the result window after performing the test, the result is considered invalid. Some causes of invalid results are not following the directions correctly or the test may have deteriorated beyond the expiration date. It is recommended that the specimen be re-tested using a new test kit.

8. Limitations of the test

1) This test is a screening test intended to provide a qualitative detection of human hemoglobin in fecal samples. A positive result suggests the presence of human hemoglobin in fecal samples. The presence of blood in stools may be due to several causes, besides colorectal bleeding, such as hemorrhoid, blood in urine or stomach irritations. 2) As with any test, the results will only be as reliable as the quality of the specimen provided will permit. 3) As with all diagnostic tests, all results must be considered with other clinical information available to the physician. So, it should be noted that a single result cannot be used to make a definitive diagnosis and that such a diagnosis should be made by the physician after all clinical and laboratory findings have been evaluated. 9. Internal Quality Control The SD BIOLINE FOB test device has Test Line and Control Line on the surface of the cassette. Both the Test Line and Control Line in result window are not visible before applying any samples. The Control Line is used for procedural control. Control line should always appear if the test procedure is performed properly and the test reagents of control line are working. 10. Performance Characteristics 1) Expected values This kit can detect human hemoglobin in fecal samples. A positive result indicates the sample probably contains human hemoglobin above the concentration of 50ng/ml in extracted solution. 2) Sensitivity and Specificity SD BIOLINE FOB Test has been compared with a leading commercial FOB rapid test. The correlation between these two systems is over 98 % as follows; Reference SD BIOLINE FOB Total Results Method Result Positive Negative A leading Commercial Positive 100 2 102 FOB test Negative 3 202 205 TOTAL RESULTS 103 204 307 In a comparison of the SD BIOLINE FOB test versus a leading commercial FOB rapid test,results gave sensitivity of 98% (100/102), a specificity of 98.5% (202/205), and a total agreement of 98.3% (302/307). 3) Precision (1) Within run precision was determined by using triplicates of 6 different specimens containing different concentrations of human blood hemoglobin. The negative and positive values were correctly identified 100% of the time. (2) Between run precision was determined by using of 6 different specimens containing different concentrations of hemoglobin in 3 different replicates with 3 different lots of test devices. Again negative and positive results were observed 100% of the time. 4) Analytical sensitivity : The limit of detection (50 ng/ml of human hemoglobin); the smallest amount of the target marker that can be precisely detected ; have been equal to a leading commercial FOB Rapid Test. 5) Analytical specificity : No cross reactivity test with following materials on SD BIOLINE FOB test; - Swine hemoglobin, Chicken hemoglobin, Bovine hemoglobin, HRPO, Bilirubin, Bovine serum albumin, Vitamin C and Sucrose. 11. Bibliography of suggested reading 1) Screening for colorectal cancer using the fecal occult blood test : a critical literature review. European Journal of Oncology Nursing Vol. 5(4) 234-243, 2001. 2) Fecal occult blood test for colorectal cancer screening. Annals of oncology 13:51-56, 2002. 3) Should organized fecal occult blood test screening be established? Annals of oncology 13:57-60, 2002. 4) Review article : fecal occult blood testing for colorectal cancer. Aliment Pharmacol Ther 1998; 12:1-10.
Date Issued : 2010. 02 25FK10-02-2, 25FK11-02-7, 25FK12-02-6

6. Procedure of the test ( Refer to figure of detailed specimen collection and test procedure.)
1) Allow test device and extracted sample to room temperature prior to testing. 2) Remove the test device from the foil pouch prior to use. Divide test device as occasion demands from multi-device, and then please seal the pouch after use to protect test device left from the humidity. 3) Shake the collection tube thoroughly to ensure proper mixing of the fecal sample with the assay diluent. 4) Loosen cap on the lower part of the sample collection tube. Hold the collection tube vertically and add 3 drops (about 120 ) into the sample well (S) of test device 5) Interpret test results at 5 ~ 10 minutes. Do not interpret test result after10 minutes. 6) If the sample is strong positive, the result can be shown within 2-3 minutes. Interpret the result upon showing of T-line and C-line. But, if the sample is very weak positive or negative sample, interpret the result at 10 minutes. A color band appearing on C-line and a clear background in the result window is considered internal quality control. However, when the fecal samples are tested, the background may appear slightly yellowish due to the original color of the fecal samples. Caution: The above interpreting time is based on reading the test results at room temperature. If your room temperature is significantly lower than 10, then the interpreting time should be properly extended to another 10 minutes. Fill up the ditch

2. Materials provided / Active ingredients of main components (50 tests/kit)

1) SD BIOLINE FOB test kit contains following items to perform the assay; (1) 10 Test multi-device individually foil pouched with a desiccant ............................ 105 pouches (2) Sample collection tubes with assay diluent (2 ml/tube) ........................................................................... 50 tubes (3) Storage and transport bags for specimen container ................................................................................... 50 bags (4) Instruction for use 2) Active ingredients of main components (1) 1 test strip includes ; Gold Conjugates (as main component) : Mouse monoclonal anti-human hemoglobin gold colloid (10.2 ), Test Line (as main component) : Mouse monoclonal anti-human hemoglobin (40.8 ), Control Line (as main component) : Goat anti- mouse IgG (20.4 ) (2) 2 ml of assay diluent includes ; Disodium hydrogen phosphate (Na 2HPO4, 5.68 mg), Bovine serum albumin (1%), Sodium azide (0.01%.), Sodium chloride (0.5M) and Tween-20 (0.5%).

3. Kit storage & stability

1) The SD BIOLINE FOB test should be stored at room temperature (2 ~ 30). 2) Do not store the test kit in direct sunlight. 3) Do not store at refrigerator. Do not freeze. 4) Do not use it beyond the expiration. 5) The shelf-life of the kits is as indicated on the outer package. Wait 5~10 min.

4. Warnings & Precaution

1) Precautions (1) This FOB test kit is intended for professional in vitro diagnostic use only. (2) For the best results, strict adherence to these instructions is required. (3) All specimens should be handled as being capable of transmitting disease. (4) Do not open or remove test devices from their individually sealed pouched until immediately before their use. (5) All reagents must be at room temperature before running the assay. (6) Do not use reagents beyond the stated expiration date marked on the package label. (7) The components in this kit have been quality control tested as standard batch unit. Do not mix components from different lot numbers. (8) The assay diluent contains low concentration of sodium azide as a preservative. Sodium azide is toxic and should be handled carefully to avoid ingestion and skin contact. (9) Patients should closely follow the specimen collection procedures. Patients should not collect samples during their menstrual period, if they have bleeding hemorrhoids, blood in the urine, or if they have strained during bowel movement. 2) Warnings (1) For in vitro diagnostic use only. Do Not Re-use test device. (2) The instruction must be followed exactly to get accurate results. Anyone performing an assay with this product must be trained in its use and must be experienced in laboratory procedures. (3) Do not eat or smoke while handling specimens. (4) Wear protective gloves while handling specimens. Wash hands thoroughly afterwards. (5) Avoid splashing or aerosol formation. (6) Clean up spills thoroughly using an appropriate disinfectant. (7) Decontaminate and dispose of all specimens, reaction kits and potentially contaminated materials, as if they were infectious waste, in a biohazard container.

Read the result

Loosen Insert and turn Cap the stick into the fecal sample 5~6 times at different site

Re-insert the Shake applicator and mix stick into the well sample collection tube

Loosen green cap the lower part of the sample collection tube

Add 3 drops into sample well

7. Interpretation of the test

1) A color band will appear in the left section of the result window to show that the test is working properly. This band is the Control Band. 2) The right section of the result window indicates the test results. If another color band appears in the right section of the result window, this band is the Test Band. Negative Result : Only one line appears. The presence of only one purple color band within the result window indicates a negative result.

Positive Result : Two lines appear. The presence of two color bands (T band and C band) within the result window, no matter which band appears first, indicates a positive result. Note: A positive result will not change once it has been established at 10 minutes. However, in order to prevent any incorrect results, the test result should not be interpreted after 10 minutes.