Home Marihuana Test

One Step-Membrane Immunoassay for Qualitative Detection of Marijuana and Its Metabolite in Human Urine

INTENDED USE 
The Marijuana test kit is an easy to use immunoassay for use in the qualitative analysis for Marijuana and its metabolite in human urine. It should not be used without proper supervision and is not intended for sale to the general public. The Marijuana provides only a preliminary screening test result; a more specific alternate chemical method must be used in order to confirm the analytical result. Gas chromatography/Mass spectrometry (GC/MS) is the preferred confirmatory method(1). Clinical considerations and professional judgment should be applied to any drug of abuse analysis, particularly when positive results are indicated(2,3).

SUMMARY AND EXPLANATION OF TEST
The Marijuana test employs a unique monoclonal antibody set to selectively identify THC metabolites in urine with a high degree of sensitivity.
Marijuana is major drug of abuse. After smoking or oral administration, the major psychoactive compound, tetrahydrocannabinol, is extensively metabolized before excretion(7). The excretion half-life of cannabinoid metabolites in urine, estimated by different techniques, ranges from one to ten days.
For an initial immunoassay result, a cutoff value of 50ng/ml total cannabinoid metabolites is required by the mandatory reference guidelines from the U.S. Department of Health and Human Services. (8) All presumptive positive results must be confirmed at a cutoff concentration of 15ng/ml of 11-nor-delta9-tetrahydrocannabinol-9-carboxylic acid.

The Marijuana test is an easy, fast, and visually read screening test that does not require instrumentation.

PRINCIPLES OF PROCEDURE
The Marijuana is a one-step membrane immuno-chromatographic test (6) based on antigen/antibody complexation and is used for the analysis of Marijuana and its metabolite present in the test sample. The assay is based on the competition between the drug or drug metabolite in the sample and a drug conjugate immobilized on a porous membrane support for limited antibody sites on the colored microspheres. Test urine is delivered into a sample site located at one end of the porous membrane. When the drug is present in the urine test sample, the drug or drug metabolite competes for the limited antibody sites on the microspheres. When an adequate amount of drug is present, attachment of the colored microspheres to the probe site on the membrane occurs. Therefore, a positive urine sample will inhibit the formation of a precipitin line at the probe area. A reference or control line with a secondary antibody reaction is added to the membrane test to indicate that the test is viable. This control line should always be present before making any test interpretation. Normally, a negative urine sample will produce two colored lines (the formation of a visible precipitin line at the test zone in addition to the control line), and a positive urine sample will show only one line (the control line).

REAGENTS
Each Marijuana test is individually sealed in a foil pouch.

Each test contains a membrane strip with a defined amount of colloidal gold conjugated with monoclonal (mouse) or polyclonal antibodies (sheep) in dry chemical form as absorbed in pad. A secondary polyclonal antibody (goat anti-mouse lgG) is used to form the reference line. Also, an antigenic drug conjugate an animal serum are adsorbed onto the membrane. The entire membrane is then dried before assembly and is used in the dry form. All necessary reagents for performing the test are preformulated and included in each individual test. No additional reagents are required. One needs only to add the test sample.

MATERIALS PROVIDED
Each Marijuana test (one cassette or one strip) individually sealed in a foil pouch
Each pouch contains:
1.One Marijuana test
2.Desiccant
3.Urine dropper (for cassette)

Material required but not supplied:

- Timer, sample container and disposable gloves.
- No other equipment or reagents are needed

STORAGE
The components of this kit are stable until the marked expiration data when stored in the sealed pouch at 2 o-30 oC (36 o-86 oF).

• The test must be brought to room temperature prior to use. Do not allow test to be subjected to below freezing or above 30 oC temperatures.
• Urine samples if not used immediately after collection, should be stored refrigerated (2-8 oC).
• For storage longer than 3 days, the samples should be frozen(< -20 oC)

WARNINGS AND PRECAUTIONS

1. Avoid cross-contamination of urine samples by using a new urine specimen cup and dropper for each urine sample.
2.If a test foil bag is torn or perforated, the test should not be used.
3.Do not use the assay beyond the expiration date as indicated on the pouch.
4.Open sealed pouch containing the test immediately prior to use. Prolonged exposure to ambient humidity will cause product deterioration.
5. Urine samples should not exhibit a strong positive test for protein since either heavy proteinuria or hematuria may adversely affect the performance of the test.
WARNING: Urine specimens and all materials coming in contact with them should be handled and disposed of as if infectious and transmitting infection. Avoid contact with skin.

ASSAY PROCEDURE (CASSETTE)

1. SPECIMEN COLLECTION AND HANDLING

The Marijuana test is formulated for use with urine specimens. Fresh urine specimens should be used, and the urine does mot require any special pretreatment. Urine samples should be collected such that testing may be performed as soon as possible, preferable during the same day as specimen collection.

The sample should be free from gross debris. Samples taking out from a refrigerated storage must be allowed to warm up to room temperature before testing.

Allow at least 30 minutes when the sample was first taken out from a refrigerated storage at 2-8 oC, give a longer time if the sample needs to be thawed out first from frozen storage.

2.TEST PROCEDURE   1)Remove the test device from its foil wrapper by tearing along the slice. 2) Using the sample dispenser, withdraw the urine sample from the specimen cup and slowly dispense 3 drops (approximately 120uL) into the circular sample well, being careful not to overfill the absorbent pad. (See A) 3)Read results within five minutes to achieve accurate specimen analysis. DO NOT INTERPRET RESULT AFTER 10 MINUTES.

3. RESULT READING

1)Positive: Only one color band appearing in the "c" region (see B)

2)Negative: One band appearing in the "c" region, the other in the "T" region. (See C)

3)Invalid: No color bands appearing in the window at all, the test result is invalid. (See D)  

ASSAY PROCEDURE (DIPSTICK) 1.SPECIMEN COLLECTION AND HANDLING       (PLEASE REFER TO THE SAME SECTION OF CASSETTE) 2.TEST PROCEDURE

1)Remove the strip from the foil wrapper or the desiccated container (bring the container to the room temperature before opening to avoid condensation of moisture in container). Label the strip with patient or control identifications.   2)Immerse the strip into the urine with the arrow end pointing toward the urine. Do not cover the urine over the MAX (maximum) line. You may leave the strip in the urine or you may take the strip out after a minimum of 15 seconds in the urine and lay the strip flatly on a non-absorptive clean surface. (See E)

3)Read the result at 5 minutes.

DO NOT INTERPRET RESULT AFTER 10 MINUTES.

3.RESULT READING

1)Positive: One pink-purple colored band appears in the CONTROL area

2)Negative: In addition the control band a clearly distinguishable pink-purple colored band also appears in the TEST area.

QUALITH CONTROL
Quality control specimens are available from commercial sources such as Bio-Rad Lyphochek urine toxicology controls which can be used for the periodic assessment of positive results obtained with Marijuana.

PERFORMANCE CHARACTERISTICS
1. SENSITIVITY
In contrast to GC/MS which detects a specific drug of drug metabolite, Marijuana immunoassay procedures can detect the parent Marijuana molecule and Marijuana-like metabolites. DINA has suggested that the screening cutoff for positive samples be 50ng/ml for Marijuana. The Marijuana has been shown to detect an average of 50 ng/ml for Marijuana in urine.

2. ACCURACY
The accuracy of our Marijuana Test was evaluated in comparison to a commercially available immunoassay. One hundred sixty (160) urine samples collected from presumed non-user volunteers and Approximately eighty (80) positive sample from clinical Iaboratory have been tested by both methods. 100% agreement on negative samples and 98% on positives.

3. SPECIFICITY
The test selectively detects cannabinoid metabolites in urine. The specificity was tested by adding the following drugs, drug metabolites, or other compounds to drug-free urine samples.

REFERENCES
1.Johansson, E, Gillespie, H.K., Halldin, M.M.J. Anal. Toxicol., 14:176-180(1990).
2.El Sohly, M.A., Jones, A.B., El Sohly, H.N.J.Anall.Toxicol., 14:277-29(1990).
3.Foltz, R.L., Sunshine, I.J.Anal. Toxicol., 14:375-378(1990).
4.Wimbish, G.H., Johnson, K.D.J. Anal. Toxicol., 14:292-295(1990)
5.Nakamura, G.R., Meeks, R.D., Stall, W.J.J. Forensic Sci., 35(4):792-796(1990).
6.Jenkins, A.J., Mills, L.C., Darwin, W.D., Huestis. M.A., Cone, E.J., Mitchell, J.M.J. Anal. Toxicol., 17:292-298(1993).
7.Hollister, L.E., Kanter, S.L., Board, R.D., Green, D.E.Res. Com.Chem.Pathol.Pharmacol., 8:579-584(1974).
8. Federal Register 53:11970-11983(1988).