BodiTech Med Inc. i-CHROMATM FOB
i-CHROMATM FOB
ImmunoAssay for Quantitative Measurement of Fecal Occult Blood (FOB) in Human feces with i-CHROMATM Reader System.
INTENDED USE
The i-CHROMATM FOB test along with i -CHROMATM Reader is a fluorescence immunoassay that measures of hemoglobin (Hb) in human feces specimens.
INTRODUCTION
Colorectal cancer is the third most common cancer in world1, with about 1 million new cases and more than 500 000 deaths per year. Screening method for colorectal cancer include the fecal occult blood (FOB) test, barium enema, sigmoidoscopy and colonoscopy2. Large randomized controlled trials have shown that FOB screening can result in decreased colorectal cancer mortality3,4. The standard FOB test uses the chemical Guaiac, which is sensitive to Hb peroxidase activity. However, the standard Guaiac-FOB test has low sensitivity for clinically significant colorectal neoplasis and has low specificity due to its non-specificity for human Hb5,6. To overcome these potential problems in immunochemical test, i-CHROMATM FOB uses the specific monoclonal antibodies against human Hb as capture and detector reagents7.
PRINCIPLE
i-CHROMATM FOB is based on fluorescence immunoassay technology. The i-CHROMATM FOB uses a sandwich immuno-detection method, such that by mixing detection buffer with human Hb specimen in sample collection tube, the fluorescence-labeled detector anti-Hb antibody in buffer binds to Hb in fecal specimen. As the sample mixture is loaded onto the sample well of the test device and migrates the nitrocellulose matrix of test strip by capillary action, the complexes of detector antibody and FOB are captured to anti-human Hb sandwich pair antibody that has been immobilized on test strip. Thus the more Hb is in fecal specimen, the more complexes are accumulated on test strip. Signal intensity of fluorescence of detector antibody reflects amount of human Hb captured and is processed from i-CHROMATM Reader to show Hb concentration in fecal specimen. The result unit of i-CHROMATM FOB is displayed as an ng/mL from i-CHROMATM Reader. The working range of i-CHROMATM FOB test system are 25 ~ 1,000 ng/mL.
* Reference Value: 50 ng/mL
MATERIALS PROVIDED
BodiTech Med Incorporated i-CHROMATM FOB
REF Catalog No. 13016
kit contains:
Test Device 25T/box
Sample Collection Tube 25 tubes/box (1.5ml/tube)
ID Chip 1 ea/box
Insert 1 ea/box
MATERIALS REQUIRED BUT NOT PROVIDED
i-CHROMATM Reader REF Catalog No. FR-203
Thermal Printer (optional)
COMPOSITION OF REAGENTS
i-CHROMATM FOB kit consists of test device and sample collection tube. Test device is individually sealed with a desiccant in aluminum pouch, and sample collection tube is packed and delivered separately from test device in a styrofoam box filled with ice pack.
• Test device contains a test strip in which murine monoclonal antibody against human Hb and rabbit IgG have been immobilized on the test and on the control line of strip, respectively.
• Sample collection tube contains fluorescence-labeled anti-human Hb (mouse monoclonal), fluorescence-labeled anti-rabbit IgG, BSA as a stabilizer, Tween 20, and sodium azide as a preservative in PBS.
WARNINGS AND PRECAUTIONS
• IVD For In Vitro Diagnostic Use.
• Carefully follow the instructions and procedures described in this insert. REF Catalog No. 13016
• Don’t use test device if its lot # does not match with ID Chip # that is inserted onto the instrument.
• i-CHROMATM FOB kit is only operational in the i-CHROMATM Reader. And tests should be applied by professionally trained staff working in certified laboratories at some remove from the patient and clinic at which the sample(s) is taken by qualified medical personnel.
• LOT Neither interchange materials from different product lots nor use beyond the expiration date. The use of medical device beyond expiration date may affect on test result.
• i-CHROMATM FOB should remain in its original sealed pouch until ready to use. Do not use the test device if the pouch is damaged or the seal is broken. Discard after single use.
• FOB test device and reader should be used away from vibration and magnetic field. During normal usage, i-CHROMATM FOB test may introduce minute vibration, which should be regarded normal.
• Use separate clean sample collection tube. Sample collection tube should be used for one specimen only. Discard after single use.
• Fecal specimens, used test device and sample collection tube are potentially infectious. Proper laboratory safety techniques, handling and disposal methods should be followed in accordance with standard procedures and relevant regulations observed by microbiological hazard materials.
• The test will be applied on a routine basis and not in emergency situations.
• Do not smoke, eat, or drink in areas in which specimens or kit reagents are handled.
• Care should be taken to avoid contamination of the end of bottle when dropping of assay diluents into sample well.
STROAGE AND STABILITY
• Store the sample collection tube in a refrigerator at 2° - 8°C. Sample collection tube is stable up to 20 months if stored in a refrigerator.
• Once removed from refrigerator, allow the sample collection tube for 30 minutes to return to room temperature before testing.
• Store test device at 4°-30°C in its sealed pouch. Test device is stable for 20 months (while in the sealed pouch) if stored at 4°-30°C.
• If stored in a refrigerator, allow a minimum of 10 minutes for the test device to reach room temperature while it is in the sealed pouch.
• Do not remove the device from the pouch until ready to use. Test device should be used immediately once opened.
• The storage and shipping of test kit should be complied as indicated in manual. However, it is remotely possible that only part of test kit is affected by stability problems.
SAMPLE COLLECTION AND PREPARATION
The test can be performed fecal specimens.
• Collect a random sample of feces in clean, dry container or receptacle, making sure to exclude.
• Loosen cap on the upper part of sample collection tubes and remove sampling 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.
• Collect random sample by using the sampling stick in the proper method, that is, insert the sampling 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.
• Re-insert the sampling stick into the sample collection tube and screw the cap tightly.
• Shake and mix well the tube vigorously to provide a good homogenous extraction and suspension.
ASSAY PROCEDURE
• Image of the test kit
window sample well
1. Set a Test Device on a dust-free clean place.2. Check/insert ID Chip onto the instrument. Make sure that the Test Device lot # matches with ID Chip #.
3. Take out one Sample Collection Tube from refrigerator and leave it at room temperature.
4. Remove the upper cap with sampling stick from the Sample Collection Tube.
5. Put the sampling stick into the fecal sample about 5 ~ 6 times at different sites and try to avoid obtaining clumps of fecal matter.
6. Return the sampling stick with feces into the Sample Collection Tube. Tighten the upper cap and then shake the tube up and down to make sure that the feces are mixed well with the buffer.
7. Take out the lower cap of Sample Collection Tube. Squeeze the tube slightly and discard the first 2 drops of mixed sample into a waste container.
8. Load next 2 drops into the well of Test Device. (Make sure of putting 2 drops to get an accurate result of the test.)
9. Leave Test Device at room temperature for 10 minutes before inserting the device into the holder of i-CHROMATM Reader*.
10. To start scanning, insert Test Device onto the holder of i-CHROMATM Reader and press “SELECT” button. Make sure direction of Test Device and push the device back all the way.
11. The instrument will automatically start to scan the Test Device immediately.
12. Read the results on the display screen of i-CHROMATM Reader.
Ø Refer to i-CHROMATM Reader Operation Manual for the complete instructions on use of the Test.
RESULT
i-CHROMATM Reader calculates FOB test results automatically and displays FOB concentration on the screen as form of ng/mL. For further information, refer to the Operation Manual for the i-CHROMATM Reader.
QUALITY CONTROL
A quality control test should be performed as a part of good testing practice, to confirm the expected QC results, to confirm the validity of the assay, and to assure the accuracy of patient results. QC specimens should also be run whenever there is any question concerning the validity of results obtained. Upon confirmation of the expected results, the test device is ready to use with patient specimens. Control standards are not provided with this test kit. If you want to perform QC of test kit, we recommend using BodiTech Med’s control reagent. For information about obtaining the controls, contact BodiTech Med Incorporat ed’s Technical Services for assistance.
i-CHROMATM FOB test system contains internal procedure control that satisfies routine quality control requirements. This internal control is performed each time a patient sample is tested. This control indicates that the test device is inserted and read properly by i-CHROMATM Reader. An invalid result from the internal control causes an error message on i-CHROMATM Reader, indicating that the test should be repeated.
LIMITATIONS OF THE PROCEDURE
• The results of i-CHROMATM FOB test should be evaluated with all clinical and laboratory data available. If FOB test results do not agree with the clinical evaluation, additional tests should be performed.
• A positive result suggests the presence of human Hb samples. The presence of blood in stools may be due to several causes, besides colorectal bleeding, such as hemorrhoid, stomach and bronchus irritation. As with any test, the results will only be as reliable as the quality of the specimen provided will permit.
• The effectiveness of the test is highly dependent on storage of kits and sample specimens at optimal conditions. All results must be considered with other clinical information available to the physician. So, a definitive diagnosis should be made after all clinical and laboratory findings have been evaluated.
• Other factors may interfere with i-CHROMA TM FOB test and may cause erroneous results. These include technical or procedural errors, as well as additional substances in fecal specimens.
PERFORMANCE CHARACTERISTICS
1. Analytical Sensitivity: i-CHROMATM FOB test system was evaluated on the limit of detection. Three different lots of Test Devices were evaluated with 10 times of each lot. Minimum detection was calculated by average of specimens (0 at value) ± 3SD. The limit of i-CHROMATM FOB test was determined to be 25 ng/mL.
2. Specificity: There was no significant interference with other bio-molecules, such as bovine Hb, swine Hb, chicken Hb, bilirubin, albumin, and vitamin C in measurement of FOB concentration.
3. Imprecision: For the intra-assay imprecision, 20 replicates were tested at each control sample. For the inter-assay imprecision, tests were conducted on 10 sequential days, with 10 runs per day with 10 replicates at each FOB concentration.
Imprecision of i-CHROMATM FOB testFOB (ng/mL) / Intra-assay Inter-assay
Mean S.D CV% Mean S.D CV%
30 / 31.0 1.6 5.2 / 30.7 2.0 6.6
50 / 51.2 3.0 5.9 / 50.0 2.9 5.8
250
700 / 250.8 18.4 7.3
695.6 32.3 4.6 / 245.7 14.8 6.0
692.0 31.0 4.8
4. Linearity: The high concentration (700 ng/mL) was diluted with the low concentration to the following final percentages; 100%, 75%, 50%, 25%, 10%, 7% and 3%. Sample was assayed in triplicate in one analytical run at each FOB level. The coefficient of linear regression was R=0.999.
5. Comparability: The FOB concentrations of 100 clinical specimens were quantified independently with i-CHROMATM FOB and Eiken OC-SENSOR Micro automatic analyzer. The test results were analyzed and their compatibilities were investigated with linear regression and correlation of coefficient (R). The value of correlation of coefficient was 0.930 between two methods.
REFERENCES
1. Ferlay J, Bray F, Pisani P, Parkin DM. GLOBOCAN 2020: Cancer incidence, motality and Prevalence worldwide. IARC CancerBase no. 5, version 2.0. Lyon, France: IARC Pr; 2004.
2. Arnold CN, Goel A, Blum HE, Boland CR. Molecular pathogenesis of colorectal cancer: implications for molecular diagnosis. Cancer 2005;104:2035-2047.
3. Mandel JS, Bond JH, Church TR, Snover DC, Bradley GM, Schuman LM, et al. Reducing mortality from colorectal cancer by scrrning for fecal occult blood. Minnesota Colon Cancer Control study. N Engl J Med 1993;328:1365-1371.
4. Kronborg O, Fenger C, Olden J, Jorgensen OD, Sondergaard O. Randomised study of screening for colorectal cancer with fecal occult blood test. Lancet 1996;384:1467-1471.
5. Hardcastle JD, Chamberlain J, Robinson MH, Moss SM, Amar SS, Balfour TW, et al. Randomised controlled trial of fecal occult blood screening for colorectal cancer. Lancet 1996;348:1472-1477.
6. Rozen P, Waked A, Vilkin A, et al. Evaluation of a desk top instrument for the automated development and immunochemical quantification of fecal occult blood. Med Sci Moint 2006;12(6):MT27-32.
7. Oh SW, Moon JD, Park SY, et al. Evaluation of fluorescence hsCRP immunoassay for point of care testing. Clin Chim Acta 2005;356:172-177
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