TEST INSTRUCTIONS: DRUG: Methamphetamine Drug Urine Test Cassette
One Step MET Test Kit For Urine
(Revised Jan. 30th, 2017)
INTENDED USE
The one-step MET (Methamphetamines) test is a simple one step immunochromatographic assay for the rapid, qualitative detection of MET and its metabolites such as oxidized, deaminated derivatives in urine. The cutoff of the test is 1000 ng/ml of MET. It is the same as the SAMHSA recommended assay cutoff.
MET test provide only a preliminary analytical result. A more specific alternative chemical method must be used in order to obtain a confirmed analytical result. Gas chromatography, mass spectrometry (GC/MS) is the preferred method. Clinical consideration and professional judgment should be applied to any drug of abuse test result, particularly when preliminary positive results are used.
EXPLANATION OF THE TEST
MET is a potent sympathominetic agent with therapeutic applications. The drug can be taken orally, injected, or inhaled. Acute higher doses lead to enhanced stimulation of the central nervous system and induce euphoria, alertness, reduced appetite, and a sense of increased energy and power. Cardiovascular responses to MET include increased blood pressure and cardiac arrhythmias. More acute responses include anxiety, paranoia, hallucinations, psychotic behavior, and eventually, depression and exhaustion. The effects of MET generally last 2-4 hours, and the drug has a half-life of 9-24 hours in the body. MET is excreted in the urine primarily as amphetamine and oxidized and deaminated derivatives. However, 10-20% of MET is excreted unchanged. Thus, the presence of the parent compound in the urine indicates MET use. MET is generally detectable in the urine for 3-5 days, depending on urine pH level.
MET test is based on the principle of the highly specific immunochemical reactions between antigens and antibodies, which are used for the analysis of specific substances in urine. Major antibodies and buffers used in MET Test Kit listed as following; Goat anti-rabbit IgG (Control Line), mouse monoclonal antibody against MET (Gold Conjugate), BSA-MET (Test Line) and phosphate buffer and tris buffer. The cutoff of the test is 1000 ng/ml of MET.
MET test cassette has a letter T and C as “Test Line” and “Control Line” in the result window. Both the “Test Line” and “Control Line” in result window are not visible before applying any samples. The “Control Line” is used for procedural control. Control line should always appear if the test procedure is performed properly and the test reagents of control line are working properly.
| MATERIALS PROVIDEDMET test kit contains the following items to perform the assay: 1. MET test device. 2. Disposable sample dropper. 3. Instructions for use.MATERIALS REQUIRED BUT NOT PROVIDED 1. Specimen collection container. 2. Clock or timer.PRECAUTIONS 1. For professional in vitro diagnostic use only.2. Avoid cross contamination of urine samples by using a new urine specimen container and dropper for each urine sample.3. Urine specimens are potentially infectious. Proper handling and disposal methods should be established according to good laboratory practices. |
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4. Do not eat or smoke while handling specimen in the laboratory.
5. MET device should remain in its original sealed pouch until ready for use.
6. Do not use the test if the pouch is damaged or the seal is broken.
7. Do not use the test kit after the expiration date.
STORAGE AND STABILITY
MET test kit should be stored at 4-30 oC in the original sealed pouch. The expiration date given was determined under normal laboratory conditions.
SPECIMEN COLLECTION AND PREPARATION
1. Fresh urine specimens do not require any special handling or pretreatment.
2. Specimens should be collected in a clean glass or plastic container.
3. If testing will not be performed immediately, specimens should be refrigerated.
4. Specimens should be brought to room temperature before testing.
5. Specimens containing precipitate may yield inconsistent test results. Such specimen must be clarified prior to assaying.
PROCEDURE OF THE TEST
1. Remove the test disk from the foil pouch, and place it on a flat, dry surface.
2. Holding the sample dropper above the test disk. Squeeze 2 drops of specimen into the sample well (See following Figures)
3. Interpret the test results at 3 to 5 minutes.
INTERPRETATION OF RESULTS
1. As the test kit begins to work, a purple band will appear at the left section of the result window to show that the Control Line is working properly.
2. The right section of the result window indicates the test results. If another purple band appears at the right section of the result window, this band is the Test Band.
NEGATIVE: TWO COLOR BANDS
The appearance of two purple bands within the result window indicates a negative test result. No MET above the cut-off level has been detected. The color of the Test band may be lighter or darker than that of the Control Band.
POSITIVE: ONE COLOR BAND
The appearance of only one purple band within the result window indicates the result is positive, i.e. the specimen contains MET at a concentration above the cut-off level.
The urine specimen should be confirmed with a more specific alternative method such as GC/MS, before a positive determination is made
INVALID:
A distinct purple band should always appear in the left section of the result window. The test is invalid if no color band forms in the left section of the result window.
Note. A very faint band in the right section of the result window, visible in 5 minutes, indicates that the amount of MET in the sample is near or below the cut-off level of the test. The urine specimen should be retested, or confirmed with a more specific alternative method such as gas chromatography/mass spectrometry, before a positive determination is made.
USER QUALITY CONTROL
Control standards are not supplied with this kit; however, it is recommended that a control be tested as good laboratory testing practice. For information on how to obtain controls, contact Technical Service. Before using a new kit with patient specimens, positive (cutoff and 25% more than cutoff level) and negative (25% below cutoff level) controls should be tested to confirm the test procedure, and to verify the tests produce the expected Q.C. results.
LIMITATIONS
1. The test is designed for use with unadulterated human urine only.
2. There is a possibility that factors such as technical or procedural errors, as well as other substances in the urine samples may interfere with the test and cause erroneous results.
3. Adulterants, such as bleach and/or alum, in urine specimens may produce erroneous results regardless of the method of analysis. If adulteration is suspected, the test should be repeated with a new sample.
4. A positive test result does not provide any indication of the level of intoxication or urinary concentration.
5. The test results read after 5 minutes may not be consistent with the original reading obtained within the 5 minutes reading period. The test must be read within 5 minutes of sample application.
6. Certain medications containing MET may produce a positive result in any chemical and immunological assay.
EXPECTED VAI.UES
MET test is a qualitative assay. The amount of drugs and metabolites present in the urine cannot be estimated by the assay. The assay results distinguish positive from negative samples. A positive result indicates the sample contains MET above the cut-off concentration.
PERFORMANCE CHARACTERISTICS AND COMPARISION STUDIES
MET test has been shown to detect an average of 1000 ng/ml or more of MET metabolites in urine. The accuracy of the MET was evaluated in comparison to a commercially available immunoassay. A total of 50 negative real patients urine sample (concentration of MET range of 0-730 ng/ml) and 50 positive real patients urine sample (concentration of MET range of 1000-3215 ng/ml) were tested by both procedures. Complete agreement was observed in 100% of the samples. All positive and part of negative urine samples were confirmed by GC/MS.
PRECISION AND REPRODUCIBILLITY STUDIES
The precision of the MET assay was determined by carrying out the test with serially spiked MET urine samples. The four concentrations, at 0 ng/ml, at –25% from the cutoff (750 ng/ml), at the cutoff (1000 ng/ml) and +25% from the cutoff (1250 ng/ml) had been tested to challenge the precision of the test device. Total 50 run at 0 ng/ml, 50 run at 750 ng/ml, 200 run at 1000 ng/ml and 50 run at 1250 ng/ml. Total 350 tests were tested. About 99.14 % of the samples containing drug concentrations at or more than 25% over the cut-off level consistently showed positive results.
The reproducibility studies were carried in three different sites. The urine samples contain 0, 1000 ng/ml and 3000 ng/ml of MET had been tested by total 360 MET test kits. The samples were tested two times in the same day, and in two different assays, each day for 20 days. This permits separate tests of between-day, between-assay and within-day which they are shown consistent results.
CUTOFF STUDIES
There are total 200 urine samples including 50 samples contain zero, 50 samples below the cutoff 750 ng/ml), 50 samples at the cutoff (1000 ng/ml) and 50 samples at above the cutoff (1250 ng/ml). All 200 urine samples were tested by both and commercially available immunoassay test kit. Complete agreement was observed as 99.33 % and the test cutoff is 1000 ng/ml of MET.
SPECIFICITY AND INTERERENCE STUDIES
The following table lists compounds that are detected by the MET test. The results are expressed in terms of the concentration required to produce a positive result.
Compound Conc. (ng/ml)
D-Amphetamine 45,000
1-Amphetamine 45,000
(+)Ephedrine 150,000
(-)Ephedrine 75,000
Methylenedioxyamphetamine 30,000
(+/-)3,4-Methylenedioxymethamphetamine 2,000
(-)-Methamphetamine 25,000
Potentially interfering chemicals such as pain medication (Acetaminophen, 20 mg/dl), protein (2000 mg/dl), glucose (2000 mg/dl), hemoglobin (500 mg/dl) and pH of 6.0,7.0 and 8.0 were supplemented to normal urine specimens devoid of MET. The test gave consistently negative results. The base line urine with 1000 ng/ml MET scored consistently positive.
REFERENCES
1. Urine Testing for Drugs of Abuse, National Institute for Drug Abuse (NIDA), Research Monograph 73, 1986.
2. Norbert W. Tietz, Texbook of Clinical Chemistry, W. B. Saunders Company, p. 1735, 1986.
