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Microsoft word - 33-2972 tcac a.doc

Instant-ViewTM TCA Urine Test (Cassette)
ONE STEP ASSAY
Dispose of all specimens and used assay materials in a proper biohazard RAPID VISUAL RESULTS
FOR QUALITATIVE IN VITRO DIAGNOSTIC USE
INTENDED USE
ASSAY PROCEDURE
This device is a one-step immunoassay intended to provide qualitative rapid Refrigerated specimens or other materials must be equilibrated to
detection of tricyclic antidepressants (TCA) at a cutoff concentration of 1,000 room temperature before testing.
ng/ml in human urine. It is for health care professional use only. Remove the test device from pouch and place it on a flat surface.
This assay provides only a preliminary result. Clinical consideration and
Label the device with specimen identification. professional judgment must be applied to any drug of abuse test result, particularly
Use the supplied dropper to add four drops (about 160 µl) of the urine in evaluating a preliminary positive result. In order to obtain a confirmed analytical
specimen to the sample well marked as “S” on the device, holding the
result, a more specific alternate chemical method is needed. Gas
Chromatography/Mass Spectroscopy (GC/MS) is the preferred confirmation
method.1
Note: If migration is not observed in 30 seconds in the results window, add one or two extra drops of urine specimen. SUMMARY AND EXPLANATION OF THE TEST
Read the test result between four (4) to seven (7) minutes after adding Tricyclic Antidepressants (TCA) are a group of antidepressant drugs that contain three fused rings in their chemical structure.2 TCA can be taken orally or by intramuscularly (IM). The progressive symptomatology of TCA includes IMPORTANT: Do not read test results after seven (7) minutes.
agitation, confusion, hallucinations, hypertonicity, seizures, and EKG changes. The half-life of TCA varies from few hours to few days. The commonly used INTERPRETATION OF RESULTS
tricyclic antidepressants are excreted with a very low percentage of unchanged POSITIVE: If only the Control line (C) appears, the test indicates a positive result.
drugs in the urine, less than 1%. Therefore, detecting the metabolites of TCA in Samples with positive results should be confirmed with a more specific method
human urine has been used for screening the abuse of TCA.3, 4 This test is able before a positive determination is made. 5
to detect amitriptyline, desipramine, imipramine and nortriptyline at a cut off Control (C) Line Test (T) Line Sample Well
PRINCIPLE OF THE PROCEDURE
This assay is a one-step lateral flow chromatographic immunoassay. The test strip includes 1) a burgundy-colored conjugate pad containing mouse anti-TCA antibodies coupled to colloidal gold (the immunogen is a blend of amitriptyline, desipramine, imipramine and nortriptyline); and 2) nitrocellulose membrane containing a Test (T) line a Control (C) line. The Test line is coated with TCA- BSA, and the Control line is coated with goat anti-mouse IgG antibody. NEGATIVE: If both the Control (C) line and Test (T) line appear, the test
This test is a competitive binding immunoassay. The TCA metabolites in the Note: A very faint line in the test region should be considered negative.
urine specimen compete with the TCA-BSA, which is coated on the nitrocellulose membrane for the limited binding sites of the anti-TCA antibodies Control (C) Line Test (T) Line Sample Well in the conjugate pad. When an adequate amount of urine specimen is applied to the sample pad of the device, the urine specimen migrates by capillary action through the test strip. If the level of TCA and/or TCA metabolites in the urine specimen is below the cutoff (1,000 ng/ml), the Test line should appear as a visible burgundy line. If the level of TCA metabolites in the urine specimen is at or above the cutoff, no Test line develops. INVALID: If no Control (C) line develops within 5 minutes, the test is invalid. In
The Control line is coated with goat anti-mouse antibody, which should bind to that case, repeat the assay with a new test device. the gold-antibody conjugate and form a burgundy color line regardless of the Control (C) Line Test (T) Line Sample Well
REAGENTS AND MATERIAL SUPPLIED
Kit contains 25 test devices, each sealed in a pouch with a dropper pipette and a desiccant.
MATERIAL REQUIRED BUT NOT PROVIDED
1.
Control (C) Line Test (T) Line Sample Well STORAGE AND STABILITY
Store the kit at room temperature 46-86°F (8-30°C). Each device may be used
until the expiration date printed on the label if it remains sealed in its foil pouch
containing a desiccant.
Do not freeze the kit and/or expose the kit to the temperature over 30°C.
QUALITY CONTROL
SPECIMEN COLLECTION
Internal Quality Control
This test contains a built-in control feature, the Control (C) line. The Each urine specimen must be collected in a clean container. presence of this Control line indicates that an adequate sample volume was Specimens may be kept at room temperature for at least 8 hours, at 2-8°C used and that the reagents migrated properly. If a C line does not form, the for up to 3 days and at -20°C or lower for prolonged storage. Do not mix test is considered invalid. In this case, review the whole procedure and repeat the testing with a new device.
PRECAUTION
External Quality Control
Users should always follow the appropriate federal, state, and local The instructions must be followed to obtain accurate results. guidelines concerning the running of external quality controls. SAMHSA6 Do not open the sealed pouch, unless ready to perform the assay. recommends that the concentration of drug(s) in positive and negative Instant-ViewTM TCA Urine Test (Cassette)
controls be approximately 25% above and below the cutoff concentration of with the Instant-ViewTM TCA Urine Test, showed a positive response at the Description Concentration
LIMITATIONS
(ng/ml)
Results obtained by this device provide only a preliminary qualitative analytical test result. A more specific alternate chemical method must be used in order to obtain a confirmed analytical result. This product is designed for testing human urine only. Adulterants such as bleach or other strong oxidizing agents may produce erroneous test results if added in the device. When suspected, collect a fresh specimen and repeat the test with a new device. Samples in which bacterial contamination is suspected should not be used. These samples may interfere with the test and cause false results. EXPECTED VALUES
This test is designed to detect TCA and/or metabolites in human urine at a cutoff concentration of 1,000 ng/ml. PERFORMANCE CHARACTERISTICS
4. Interference
To evaluate the possible interference of structurally unrelated compounds 1. Accuracy
One hundred (100) human urine specimens were used for the accuracy study with the Instant-View TCA Urine Test, the following analytes, usually on the device. Of the 100 specimens, 50 were negative and the other 50 were found in urine and commonly prescribed therapeutic drugs, were spiked in positive. Within the 50 negative specimens, 40 were without TCA, and 10 drug-free urine pools, as well as TCA positive (spiked with TCA to the level were 25% below cutoff (764 ng/ml). Within the 50 positive specimens, 10 of 1,000 ng/ml) urine pools accordingly, and then tested with the Instant- were at the cutoff (1024 ng/ml), 10 were 25% above cutoff (1224 ng/ml), 18 ViewTM TCA Urine Test. No significant interference with either negative or were 50% above cutoff (1425 ng/ml), and 12 were positive (1825 ng/ml). The positive results was observed at the concentrations listed below: specimens were calibrated with GC/MS method. Compounds tested and found not to interfere with the
An in-house accuracy evaluation was performed. Specimens were blindly test at 1.0 mg/ml concentration in urine
labeled. The results obtained from the Instant-ViewTM TCA Urine Test were 48 negative and 52 positive. The negative results agreed 96% (48/50) with the GC/MS method. The positive results agreed 100% (50/50). The two (2) discrepancies were within the range of 25% below the cutoff level. The overall correlation of the results was 98% with GC/MS results. Benzoylecgonine Phenylpropanalamine Therefore, the accuracy of the Instant-ViewTM TCA Urine Test is 98%. 2. Reproducibility
Biological analytes tested and found no interference
Clinical Evaluation
with the test at the concentrations listed
One hundred (100) GC/MS-verified human urine specimens were used in this Biological Analytes
Concentration
study. Of the specimens, 40 were without TCA, 10 were 25% below the cutoff, 10 were at the cutoff level, 10 were 25% above the cutoff, 18 were 50% above the cutoff, and 12 were positive. The Instant-View TCA Urine Test devices were evaluated at three Physician’s Office Laboratories (POL) and one reference laboratory. Evaluations were carried out by personnel with diverse educational backgrounds and working experiences. Three (3) discrepancies observed were within the range of 25% below the cutoff level, and all other results obtained were as expected. The results from the four evaluation sites agreed 99% with each other and 97% with GC/MS data, indicating a high reproducibility of the device. REFERENCES
FDA Guidance for Labeling Urine Drugs of Abuse Screening Testing, Kshit Dorland’s Illustrated Medical Dictionary, 26th Edition, W.B. Saunders Company, Philadelphia, PA, pp89, 1981. 4Urine Testing for Drugs of Abuse, National Institute on Drug Abuse (NIDA): Research Monograph 73, 1986. Wilson, John, Abused Drugs II, a Laboratory Pocket Guide. AACC Press., In-house Evaluation
Baselt, R.C. Disposition of Toxic Drugs and Chemicals in Man, 4th ED., The precision of the device was determined by replicate assays with three Biomedical Publ., Davis, CA; pp 35-39, 215-218, 392-395, 562-564, 1995. different lots. Specimens used in this study were TCA-spiked pooled urine at Urine Testing for Drugs of Abuse, National Institute on Drug Abuse (NIDA): six levels, the same as those in accuracy study. The devices were tested for five consecutive days five times each, for a total of 25 assays for each control. Department of Health and Human Services, Mandatory Guidelines for Federal Workplace Drug Testing Programs, Fed. Register. 53 (69): 11970 (1988). The results obtained indicate 100% precision for the replicate within each lot and no appreciable inter-lot variation across the three different lots of devices. 3. Cross-Reactivity
A study was conducted with the Instant-View TCA Urine Test to evaluate the cross-reactivity of compounds structurally related to TCA. The following compounds, when spiked into known drug-free urine pools and then tested

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