One step Cocaine Strip

　

INTENDED USE

One Step Cocaine Strip assay is a rapid, qualitative, competitive binding immunoassay for the determination of cocaine and its metabolites in urine at or above the cutoff level of 300 ng/ml.  One Step Cocaine Strip is not intended to monitor drug levels, but only to screen urine for the presence of cocaine and its metabolites.

Note: The test provides only preliminary data which should be confirmed by other methods such as gas chromatography/mass spectrometry (GC/MS).  Clinical considerations and professional judgment should be applied to any drug of abuse test result, particularly when preliminary positive results are indicated. 

　

SUMMARY AND EXPLANATION OF THE TEST

The One Step Cocaine Strip is an easy, fast, and visually read competitive binding immunoassay method for screening without the need for instrumentation to arrive at a determination.  The method employs a unique mixture of monoclonal and polyclonal antibodies to selectively identify Cocaine in test samples with a high degree of sensitivity.

Cocaine is an alkaloid present in Coca leaves (Erythyroxine coca).  Its pharmacological properties, such as stimulating and euphoric effects, have been known for centuries.5  Cocaine has been used medicinally as a local anesthetic agent, but its addictive properties have minimized its modern daily use.6

Cocaine is most often self-administered by IV (intra venous) injection, nasal insufflation (snorting), and inhalation of vapor from the heating of the free base (smoking).  The availability of "crack" (a street form of the free base) has increased the use by this latter route.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 benzoyl ecgonine, a hydrolytic degradation product, persists in urine at a detectable level for 48 hours.8

Immunoassay testing has been developed for the determination of benzoyl ecgonine in urine at concentrations of 300 ng/ml or greater, the sensitivity set by the National Institute on Drug Abuse for the screening method of drugs of abuse.9

PRINCIPLE of the test

The One Step Cocaine Strip consists of a chromatographic absorbent device in which the drug or drug metabolites in the sample compete with a drug conjugate immobilized on a porous membrane for limited antibody sites.  As the test sample flows up through the absorbent device, the free drug in the specimen competes with immobilized antigen conjugate in the test zone by binding to the antibody-dye conjugate forming an antibody-antigen complex and preventing the formation of a rose-pink color band when the drug is at or above the detection level of 300 ng/ml. 

In the case where free drug in the sample is below the detection level of 300 ng/ml, antibody-dye conjugate is free to bind to the immobilized antigen in the test zone, producing a rose-pink color band. Furthermore, unbound dye conjugate binds to the reagent in the control zone, producing a rose-pink color band, demonstrating that the reagents and device are functioning correctly.

A NEGATIVE specimen produces two distinct color bands, one in the test zone and one in the control zone.

A POSITIVE specimen produces one color band in the control zone only.

　

REAGENTS AND MATERIALS Provided

1.     Test Devices      The test device contains membrane-immobilized reagents in a protein matrix containing sodium azide. 

2.     Urine Cups (optional)           

3.     Test Instructions                   

 

MATERIALS REQUIRED BUT NOT PROVIDED

1.     Clock or timer.

WARNINGS AND PRECAUTIONS

1.     For in vitro diagnostic and professional use only.

2.     Do not use the test strip beyond the expiration date.

3.     Urine specimens may be infectious; properly handle and dispose of all used reaction devices in the biohazard container.

4.     Visually inspect the foil package to insure it is intact.  If the package is not intact, discard the device.

 

STORAGE and stability

Store test kit below 28°C; do not freeze.  Refer to the expiration date for stability.

 

SAMPLE COLLECTION AND PREPARATION

The sample must be collected in clean, dry container, either plastic or glass, without any preservatives.  Urine specimens may be refrigerated (2°-8°C) and stored up to 48 hours, or frozen (-20°C or below) prior to assaying.  If samples are refrigerated or frozen they should be allowed to come to room temperature before testing.  Urine samples exhibiting visible precipitates should be filtered, centrifuged or allowed to settle so that clear aliquots can be obtained for testing.

 

TEST PROCEDURE

1.     Bring test components and urine sample to room temperature (15°-28ºC).

2.     Do not break the seal of the pouch until ready to begin testing.

3.     Open the foil pouch at the notch and remove the test dipstick.  Place the dipstick into the urine sample. Do not allow the urine level to go above the maximum-level line indicated by the arrows.

4.     Read results at 5 minutes.

        IMPORTANT:  The result must be interpreted at five minutes.  Waiting more than five minutes may cause the reading to be inaccurate.  To avoid confusion, discard the test device after interpreting the result.

 

 INTERPRETATION OF RESULTStc "INTERPRETATION OF RESULTS"

　

 1.    Positive.  One rose-pink color band appears in the Control Zone ("C").  A positive result indicates that benzoyl ecgonine levels are at or above 300 ng/ml.

2.   Negative.  Two rose-pink color bands appear-- one in the Control Zone ("C") and one in the Test Zone ("T").  A negative result indicates that benzoyl ecgonine levels are below 300  ng/ml.

3.   Invalid.  No rose-pink color bands appear, or a band appears in the Test Zone ("T") but not in the Control Zone ("C").  An invalid result may be due to improper testing procedures or  deterioration of the kit components.  Repeat the assay sequence using a new device.

Note: There is no meaning attributed to line color intensity or width.

 

　

　

　

　

　

QUALITY CONTROL

An internal procedure control has been incorporated into the test to ensure proper kit performance and reliability.

The use of an external control is recommended to verify proper kit performance.  Quality control samples should be tested according to quality control requirements established by the testing laboratory.

 

LIMITATIONS OF THE TEST

1.     This product is designed for use with human urine only.

2.     Although the test is very accurate, there is a possibility that false results will occur due to the presence of interfering substances in the urine and/or factors beyond the control of the manufacturer, e.g. technical or procedural errors associated with the testing.

3.     The test is a qualitative screening assay and is not for determining quantitative concentration levels or the level of intoxication.

4.     Adulterants such as bleach or other strong oxidizing agents, when added to urine specimens, may produce erroneous test results regardless of the analysis method used.  If adulteration is suspected, obtain another urine specimen and retest.

Performance Characteristics

1.     Sensitivity.  The OneStep Cocaine Strip detects cocaine and the major metabolites of cocaine in urine at concentrations equal to or greater than 300 ng/ml, which is suggested by NIDA for the immunoassay method.

2.     Specificity.  A study was conducted with the OneStep Cocaine Strip to determine the cross-reactivity of non-cocaine related compounds with the test at concentrations much higher than normally found in the urine of people using or abusing them.  No cross-reactivity was detected with the substances listed in Table I.

        A separate study was conducted to determine the cross-reactivity of cocaine related compounds with the test.  Substances listed in Table II produced results approximately equivalent to the cut-off level for cocaine.

 

Acetaminophen Actylsalicylic Acid Amikacin Amitriptyline Ampicillin Arterenol Aspartame Atropine Sulfate Benzoic Acid Benzoylecgonine HCl Caffeine Chlorpheniramine Chlorpropmazine HCl Cimetidine Codeine Deoxyephedrine Dextromethorphan Diazepam Diethylpropion

Dephenylhydantoin Doxylamine Ecgonine HCl Ecgonine Methyl Ester Glucose Histamine Indomathacin Ketoprofen Levorphanol D -9-THC 11-nor-D  -9-carboxy-THC-9-COOH Meperidine Methylphenidate Methadone Methaqualone Morp. Glucuronide Morphine Sulfate Oxazepam Oxycodone

Pendimetrazine Penicillin G Pentobarbital d-Propoxyphene Hydrochlorothiazide Propanol Phencyclidine Phenobarbital Phentermine Phenylpropanolamine L-Phenylephrine Quinine Ranitidine Sodium Salicylate Tryptophan Tetrahydrozoline Theophylline Thioridazine Trifluoperazine

 

Table-II:    Concentration of cocaine-related compounds showing a positive response approximately equivalent to the cocaine cutoff level set for the test.

        

        Compound/Concentration in ng/ml

Cocaine	300
Benzoyl Ecgonine	300
Isoxuprine	1500

 

 

3. Accuracy:  The accuracy of the OneStep Cocaine Strip was first tested in urine samples in-house and subsequently in a clinical trial of urine samples submitted to a NIDA certified laboratory. In both cases the laboratories used EMIT II as their screening procedure. All positive samples by either screening method were confirmed by GC/MS. The data was combined and the results are summarized as follows:

 

                                     Syva EMIT II Positive         Syva EMIT II Negative

        Strip Positive                       194                               1

        Strip Negative                        0                               212

       

When compared to Emmit II the relative sensitivity was 100%. The relative specificity was 99.49%.  The concordance of the combined date with respect to Emit II  was 99.75%.

 

   4. Precision:  The precision was determined by replicate assays of three different patient urine samples with kits from three different production lots.  The resultant data indicated 100% precision for the duplicates within each lot and no appreciable inter lot variation when testing both positive and negative spiked samples across three (3) different lots of devices.

 

       

        bibliography

　

1.         D.W. Hoyt et al.  J. Am. Med. Assoc., Vol. 258 (1987), pp. 504-509.

2.         R.R. MacGregor, J.S. Fowler, and A.P. Wolf.  J. of Chromatography, Vol. 590 (1992), pp. 354-58.

3.         Urine Testing for Drugs of Abuse, NIDA Research Monograph, 73, (1986).

4.         E.J. Cone et al.  J. of Forensic Sciences, Vol. 34, No. 1 (1989), pp. 15-31.

5.         B. Holmstedt and A. Fredga.  J. of Ethnopharmacology, Vol. 3 (1981), pp. 113-47.

6.         FDA Guidence for labeling Urine Drugs of Abuse Screening Testing, Kshitij Mohan, 7/21/1987.

7.         A.R. Jeffcoat et al.  Drug Metabolism and Disposition, Vol. 17, No. 2 (1989), pp. 153-59.

8.         R.C. Baselt and R. Chang.  J. Anal. Toxicol., Vol. 11 (1987), pp. 81-82.

9.         Department of Health and Human Services, Federal Register, 53(69)  pp. 11970-89 (1988).

10.      E.J. Cone, D. Yousefnejad, and S.L. Dickerson.  J. of Forensic Sciences, Vol. 35, No. 4 (1990), pp. 786-91.

11.      T. Inaba and J. Can.  Physiol. Pharmacol., Vol. 67 (1989), pp. 1154-57.

12.      Cody, J.T., and Schwarzhoff, R., J. Anal. Toxicol., 17: 2630 (1993).

13.      Dasgupta, A., Saldana, S., Kinnaman, G., smith, M., and Johansen, K., Clin. Chem., 39(1): 104-108 (1993