ACCU-TELL BZD URINE TEST (CE)

Declaration: As with all diagnostic tests, a definitive clinical diagnosis should not be based on the result of a single test, but should only be made by the physician after all clinical and laboratory findings have been evaluated.

 

CATALOG

INTENDED USE

                                                                                PERFORMANCE CHARACTERISTICS

Accu-Tell ^® Rapid BZD Urine Test is a lateral flow, rapid immunoassay for the qualitative detection of benzodiazepines in human urine at a cut-off of 300 ng/mL. This product is used to obtain a visual, qualitative result and is intended for professional use. The assay should not be used without proper supervision and is not intended for over the counter sale to lay persons.

This assay provides only a preliminary analytical test result. A more specific alternative chemical method must be used in order to obtain a confirmed analytical result. Gas chromatography/mass spectrometry (GC/MS) has been established as the preferred con-firmatory method by the National Institute on Drug Abuse (NIDA). Clinical consideration and professional judgment should be applied to any drug of abuse test result, particularly when preliminary positive results are indicated.

Benzodiazepines are the most widely used anxialytic drugs. They are used extensively as anti-anxiety agents, hypnotics, muscle relaxants and anti-convulsants. They are taken orally or sometimes by injection. Benzodiazepines are metabolized in the liver. Some metabolites of benzodiazepines also exhibit pharmacological activities. Benzodiazepines and metabolites are excreted in the urine. Their use can result in drowsiness, confusion. Benzodiazepines potentiate alcohol and other CNS depressants. Psychological and physical dependence on benzodi-azepines can develop if high doses of the drug are given over a prolonged period. 6,7

Urine based screening tests for drugs of abuse range from simple immunoassay tests to complex analytical procedures. The speed and sensitivity of immunoassays have made them the most widely accepted method for screening urine for drugs of abuse. Rapid BZO Test Cassette is based on the principle of the highly specific immunochemical reactions of antigens and antibodies which are used for the analysis of specific compounds in biological fluids. This test is a rapid, visual, competitive immunoassay that can be used for the qualitative detection of benzodiazepines in human urine at 300 ng/ml cut-off concentration.

Accu-Tell ^® Rapid BZD Urine Test is a rapid immunoassay in which a chemically labeled drug (drug conjugate) competes with the drug which may be present in urine for limited antibody binding sites. The test device contains a membrane strip which was pre-coated with drug conjugate on the test band. A colored anti-benzodiazepines monoclonal antibody-colloidal gold conjugate pad is placed at the right end of the membrane. In the absence of drug in the urine, the solution of the colored antibody-colloidal gold conjugate and urine moves upward, chro-matographically by capillary action, across the membrane. This solution then migrates to the immobilized drug conjugate zone on the test band region. The colored antibody-colloidal gold conjugate then attaches to the drug conjugate to form a visible line as the antibody complexes with the drug conjugate. Therefore, the formation of a visible precipitant in the test zone occurs, when the test urine is negative for the drug. When the drug is present in the urine, the drug/metabolite antigen competes with the drug conjugate on the test band region for limited antibody sites on the anti-benzodiazepines monoclonal antibody-colloidal gold conjugate. When a sufficient concentration of drug is present, it will fill the limited antibody binding sites. This will prevent attachment of the colored antibody-colloidal gold conjugate to the drug conjugate zone on the test band region. Therefore, absence of the color band on the test region indicates a positive result.

A control band with a different antigen/antibody reaction is also added to the immunochromatographic membrane strip at the control region (C) to indicate that the test has performed properly. This control line should always appear, regardless of the presence of drug and metabolite. This means that negative urine will produce two colored bands, and positive urine will produce only one band. The presence of this colored band in the control region also serves as 1) verification that sufficient volume has been added, and 2) that proper flow was obtained.

The test kit is to be stored refrigerated or at room temperature 2-30°C (36-86 ⁰F) in the sealed pouch for the duration of the shelf life.

1.  For in vitro diagnostic use only.

2.  Urine specimens may be potentially infectious. Proper handling and disposal methods should be established.

3.  Avoid cross-contamination of urine samples by using a new specimen collection container and specimen pipette for each urine sample.

Accu-Tell ^® Rapid BZD Urine Test is formulated for use with urine specimens. Fresh urine does not require any special handling or pretreatment. Urine samples should be collected such that testing can be performed as soon as possible after the specimen collection, preferably during the same day. The specimen may be refrigerated at 2-8°C for 2 days, or frozen at -20°C for a longer period of time. Specimens that have been refrigerated must be equilibrated to room temperature prior to testing. Specimens previously frozen must be thawed, equilibrated to room temperature, and mixed thoroughly prior to testing.

Note: Urine specimens and all materials coming in contact with them should be handled and disposed of as if capable of transmitting infection. Avoid contact with skin by wearing gloves and proper laboratory attire.

Review “Specimen Collection“ instructions. Test cassette/stip, patient’s samples, and controls should be brought to room temperature (20-30°C) prior to testing. Do not open pouches until ready to perform the assay.

1.  Remove the test cassette from its protective pouch (bring the device to room temperature before opening the pouch to avoid condensation of moisture on the membrane). Label the device with patient or control identification.

2.  Draw the urine sample to the line marked on the pipette. Dispense 3 drops (or 120 μl) into the sample well. Use a separate pipette and device for each sample or control.

3.  Read result between 3 to 8 minutes after the addition of sample. Do not read result after 8 minutes.

1.  Bring the pouch to room temperature before opening it. Remove the test strip from the sealed pouch and use it as soon as possible.

2.  With arrows pointing toward the urine specimen, immerse the test strip vertically in the urine specimen for at least 5 seconds. Do not pass the maximum line on the test strip when immersing the strip.

3.  Place the test strip on a non-absorbent flat surface, start the timer and wait for the red line(s) to appear. The result should be read between 3 to 8 minutes after the addition of sample. Do not interpret the result after 8 minutes.

Negative: Two colored lines appear in the viewing window. The line in the test region (T) is the drug probe line; the line on the control region (C) is the control line, which is used to indicate proper performance of the device. The color intensity of the test line may be weaker or stronger than that of the control line.

Positive: Only one colored line appears in the control region (C). The absence of a test line indicates a positive result.

Invalid: No line appears in the control region. Under no circumstances should a positive sample be identified until the control line forms in the viewing area. If the control line does not form, the test result is inconclusive and should be repeated.

Note: A very faint line in the test region indicates that the barbiturate in the sample is near the cut-off level of the test. These samples should be re-tested or confirmed with a more specific method before a positive determination is made.

1.  The assay is designed for use with human urine only.

2.  A positive result with any of the tests indicates the presence of a drug/metabolite only, and does not indicate or measure intoxication.

3.  There is a possibility that technical or procedural errors as well as other substances and factors not listed may interfere with the test and cause false results.

4.  If it is suspected that the samples have been mislabeled or tampered with, a new specimen should be collected and the test should be repeated.

Good laboratory practice recommends the use of control materials to ensure proper kit performance. Quality control specimens are available from commercial sources. When testing the positive and negative controls, use the same assay procedure as with a urine specimen

The accuracy of Accu-Tell ^® Rapid BZD Urine Test was evaluated in comparison to a commercially available immuno-assay at a cut-off of 300 ng/mL for benzodiazepines (Oxazepam). 114 urine samples, collected from presumed non-user volunteers, have been tested by both procedures with 100% agreement.

The reproducibility of Accu-Tell ^® Rapid BZD Urine Test was evaluated at 4 different sites using blind controls. Of 60 samples without oxacepam all were determined negatives. Of 60 samples with oxacepam concentration of 600 ng/mL, all were determined positive.

The precision of Accu-Tell ^® Rapid BZD Urine Test was determined by conducting the test with blind spiked controls. The control at 600 ng/mL should give a positive result.

Concentration (ng/mL)	No Tested	Correct Result	% Correct Result
0	50	50	100
600	50	50	100

The specificity for Accu-Tell ^® Rapid BZD Urine Test was tested by adding various drugs, drug metabolites, and other compounds that are likely to be present in urine. All compounds were prepared in drug-free normal human urine.

The following structurally related compounds produced positive results when tested at levels equal to or greater than the concentrations listed below.

Compound	Concentration (μg/mL)
Oxacepam	0,30
Alprazolam	1,50
Bromazepam	0,80
Chlordiazetoxide	3,00
Clobazam	2,00
Clonazepam	5,00
Clorazepam	1,00
Delorazepan	6,00
Diazepam	1,50
Estazolam	3,00
Flunitrazepam	10,00
Flurazepam	3,00
Lorazepam	15,00
Lormetazepam	10,00
Medazepam	20,00
Nitrazepam	10,00
Nordiazepam	1,00
Prazepam	10,00
Temazepam	1,50
Triazolam	15,00

The following compounds were found not to cross-react when tested at concentrations up to 100 μg/mL:

Acetaminophen, Acetone, Albumin, Amitriptyline, Ampicillin, Aspartame, Aspirin, Atropine, Benzocaine, Benzoylecgonine, Bilirubin, Caffeine, (+)-Chlorpheniramine, (+/-)-Chlorpheniramine, Chlorpromarine, Creatine, Desoxyephetrine, Dexbrompheniramine, Dexbromethorphan, 4-Di-methylaminoantipyrine, Dopamine, Doxylamine (+/-)-Ephedrine, (-)-Ephedrine, (+)-Epinephrine, Erytromycin, Ethanol, Furosemide, Glucose, Guaiacol Glycerylether, Hemoglobin, Hydromorphone, Hydro-codone, Hydroxytyramine, Imipramine, (+/-)-Isoproterenol, Lidocaine, Naloxane, Naltrexone Naphthalene acetic acid, (+)-Naproxen, (+/-)-Norephedrine, 11-nor–Δ9-THC-9-carboxylic acid, Oxalic Acid, Oxycodone, Penicillin-G, Pentamine, Pentobarbital, Pheniramine, Phenobarbital, Phenothiazine, L-Phenyl-ephrine, D-Pro-poxyphene, Quinidine, Riboflavin, Secobarbital, Sodium Chloride, Sulindac, Thio-ridazine, Trifluorperazine, Tyramine, Vitamin C

1.  Baselt, R.C. Disposition of Toxic Drugs and Chemicals in Man, Biomedical Publications, 1982.

2.  Urine Testing for Drugs of Abuse. National Institute on Drug Abuse (NIDA), Research Monograph 73, 1986.

3.  Fed. Register, Department of Health and Human Services, Mandatory Guidelines for Federal Workplace Drug Testing Programs, 53, 69, 11970, 1988.

4.  McBay, A.J. Clin. Chem. 33, 33B-40B, 1987.

5.  Gilman, A.G., & Goodman, L.S. The Pharmacological Basis of Therapeutics, eds. MacMillan Publishing, New York, NY, 1980.

6.  Greenblatt, D.J., Shader, R.I.: Benzodiazepines in Clinical Practice. New York, Raven Press, 1974.

7.  Harvey, R.A. Champe, P.C.: Lippincott’s illustrated Reviews. Pharmacology, 91-95, 1992.

 

The above information is just for reference. The actual technical specifications are subject to the insert provided with the product.