Cocaine Home Test

For Professional and in vitro Diagnostic use only. One step-Membrane Immunoassay for Qualitative Detection of Cocaine / Benzoylecgonine and Its Metabolite in Human Urine Detection of Metbamphetamine and Its Metabolite in Human Urine

INTENDED USE 
The Cocaine test kit is an easy to use immunoassay intended for use in the qualitative analysis for Cocaine 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 Cocaine test kit provides only a preliminary screening test result; a more specific alternate chemical method must be used in order to confirm the analytical results. 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 method employs unique monoclonal antibodies to selectively identify benzoylecgonine in test samples with a high degree of sensitivity. Cocaine is an alkaloid present in coca leaves ( Erythroxylon coca ) and its pharmacological properties, such as stimulating and euphoric effects have been known for many centuries.(5) Cocaine has been used medicinally as a local anesthetic agent; however due to its proven addictive properties, modern day medicinal use is avoided.(6)

Cocaine is most often self-administered by IV injection, nasal insufflation (snorting) or vapor inhalation of heated free base (smoking or "freebasing"). The availability of "crack" (a street form of the free base) has caused vapor inhalation to become much more common. (7)

Elimination of cocaine is predominantly controlled by its biotransformation.  Very low concentrations of cocaine are detected in urine during the initial several hours and benzoylectgonine, a hydrolytic degradation product, persists in urine at a detectable level for forty-eight hours. (8)

Immunoassay testing has been developed for the determination of benzoylecgonine in the urine at the concentration of 300ng/ml as set by the National Institute on Drug Abuse. (9)

The Cocaine / benzoylecgonine test kit is an easy, fast, and visually read screening method without the need for instrumentation.

PRINCIPLES OF PROCEDURE
The Cocaine test kit is a one-step membrane immuno-chromatographic test (6) based on antigen/antibody complexation and is used for the analysis of cocaine 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 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 Cocaine test kit is individually sealed in a foil pouch.

Each test kit 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 performulated and included in each individual test. No additional reagents are required. One needs only to add the test sample.

MATERIALS PROVIDED
Each Cocaine test (one cassette or one strip) individually sealed in a foil pouch.
Each pouch contains:
1.One Cocaine 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 date 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 o-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 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 Cocaine test kit is formulated for use with urine specimens. Fresh urine specimens should be used, and the urine does not 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 from when the sample was first taken out from a refrigerated storage at 2 o-8 oC; give 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. 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.

3) Inconclusive: If there is no distinct pink-purple colored band visible both in the TEST area and CONTROL area the test is inconclusive. It is recommended that in this case test be repeated or a fresh specimen be obtained and tested 48 hour later.  

LIMITATIONS OF PROCEDURE
The assay is designed for use with human urine only. A positive result from the cocaine test only indicates the presence of cocaine, or its metabolite and does not indicate or measure intoxication. There is the possibility that other substances and/or factors not listed (e.g. technical or procedural errors) may interfere with the test and cause false results. See SPECIFICITY for lists of substances that do not interfere with test performance.

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 the Cocaine test kit.

PERFORMANCE CHARACTERISTICS
1. SENSITIVITY
In contrast to GC/MS which detects a specific drug of drug metabolite.

The Cocaine test kit immunoassay procedures can detect the parent Cocaine molecule and Cocaine-like metabolites. NIDA has suggested that the screening cutoff for positive samples be 300 ng/ml for Cocaine. The Cocaine test kit has been shown to detect an average of 300 ng/ml for Cocaine in urine.

2. ACCURACY
A study was performed using positive and negative urine specimens assayed with both The Cocaine / Benzoylecgonine test kit and a commercially available test. Confirmation of results was obtained by GC/Mass.

Commercially Available Test (GC/Mass) 

The data demonstrates the excellent correlation between the Cocaine / Benzoylecgonine test kit and another commercially available test. The clinical significance of the two tests is comparable. The precision was determined using replicate assays of samples from three patients with kits from three different lots. Each test of a patient’s urine sample was run demonstrates 100% precision for the duplicates of each sample and 100% precision using the test kits from different lots.

3. SPECIFICITY
The Cocaine / Benzoylecgonine test kit can specifically detect benzoylecgonine in urine. 
There is no interference by the following substances at a 100ug/ml concentration in urine:                       

REFERENCES
1.Hoyt.D.W.,Finnigan.R.E.,Nee.T., Shults, T.F.,Bulter, T.J.JAMA 258 : 504-509 (1987).
2.MacGregor, R.R., Fowler, J.S., Wolf, A.P.J. Chromatogr. 590 : 354-358 (1992).
3.Cone, E.J.,Yousefnejad, D., Dickerson, S.L., J. Forensic. Sci., 35 :786-791 (1990).
4.Holmstedt, B., Fredga, A.J. Ethnotharmacol. 3 : 113-1157 (1981).
5.Inaba, T.Can. J. Physol. Pharmacol., 67 : 1154-1157 (1989).
6.Jeffcoat, A.R.,Perez-Reyes,M.,HILL,J.M.,Sadler, B.M.,Cook, E.E., Drug Metab. Dispos.Bilo. Fate Chem. 17 : 153-159 (1989).
7.Baselt, R.C., Chang, R.J. Anal. Toxicol., 11 : 81-82 (1987).
8.Federal Register, 53 (69) : 11970-11989 (1988).