SCREENING TEST FOR METAMPHETAMINE AND TETRAHYDROCANNABINOL IN URINE using
SD Bioline MET/THC Rapid Test (FOR SCREENING DRUG TESTING LABORATORY)
I.
PRINCIPLES AND BASES OF TESTING
A. BASIS FOR TESTING OF URINE
• The effects of methamphetamine generally last 2-4 hours, and the drug has a half-life of 9-24 hours in the
body. Methamphetamine is excreted in the urine primarily as amphetamine and oxidized and deaminated
derivatives. However, 10-20% of methamphetamine is excreted in an unchanged form. Thus, the presence of
the parent compound in the urine indicates methamphetamine use. Methamphetamine is generally
detectable in the urine for 3-5 days after ingestion, depending on urine pH level.
• THC (Δ9-tetrahydrocannabinol) is the primary active ingredient in cannabinoids (marijuana). The peak effect
of smoking THC occurs in 20-30 minutes and has a duration of 90-120 minutes after one cigarette. Elevated
levels of the urinary metabolites of THC are found within hours of exposure and remain detectable for 3-10
days after smoking. The main metabolite excreted in the urine is Δ9-THC-COOH.
B. BASIS FOR CUT-OFF VALUES FOR SCREENING TEST
Cut-off values of 1,000ng/ml for methamphetamine and 50ng/ml for Δ 9-THC-COOH have been established as a
guideline for screening results by the National Institute on Drug Abuse (NIDA).
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A sample containing methamphetamine (test band M) at 1,000ng/ml or greater is considered positive
A sample containing Δ9-THC-COOH (test band T) at 50ng/ml or greater is considered positive for THC.
Samples containing lower concentrations of these agents are considered negative.
C. TEST PRINCIPLE
The Bioline MET/THC test relies on competition for antibody binding between drug-protein conjugates and drugs
that may be present in the urine sample. It contains a membrane strip, which has two test band regions that are
pre-coated with drug-protein conjugates. A gold pad containing mouse monoclonal drug antibodies conjugated
to colloid gold particles is placed between the sample pad and the membrane. The antibodies on the test strip
allow for selective identification of methamphetamine and Δ9-THC-COOH in urine with a high degree of sensitivity.
During testing, a urine specimen migrates upward by capillary action.
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If a drug is present in the urine specimen below its cut-off concentration, or if no drug is present, the binding
sites of the respective antibody conjugated gold particles in the test device will not be saturated. The antibody
conjugated gold particles will then be captured by their respective immobilized drug-protein conjugate, and
a colored line will appear in the associated test region. The colored line will not form in the test region if the
drug level is at or above its cut-off concentration because it will saturate all the binding sites of the drug
antibodies.
A drug-positive urine specimen will not generate a colored line in the respective test region due to drug
competition, while a drug-negative urine specimen will generate a line in the respective test band region due
to the absence of drug competition.
Regardless of whether a test band appears in either test region (M or T), a control band should be present in
the control region (C) to confirm the viability of the test. The control band is immobilized with goat anti-mouse
immunoglobulin, and will therefore capture any monoclonal antibody-gold conjugate that passes through the
region, producing a colored band in the control region (C). This functions as a procedural control. The control
band should always appear if the test procedure is performed properly and the test reagents of the control
line are working.
II.
SPECIMEN COLLECTION AND STORAGE
• Urine specimens should be collected in a clean glass or plastic container. Fresh urine specimens do not
require any special handling or pretreatment.
• If specimens are not immediately tested, they should be refrigerated at 2-8°C or frozen. Specimens
should be brought to room temperature (15-30°C) before testing.
• Specimens containing precipitate may yield inconsistent test results. Such specimens must be clarified
by centrifuging or allowing to settle prior to assaying.
• For prolonged storage, specimens may be frozen and stored at a temperature below -20°C. Frozen
specimens should be completely thawed, thoroughly mixed and brought to room temperature (15-30°C)
prior to testing.
III.
MATERIALS AND ACTIVE INGREDIENTS OF MAIN COMPONENTS
One test kit contains:
• 25 Test devices with desiccant in individual foil pouches
• 25 Disposable urine droppers
• 1 Instructions for use
Active ingredients of test strips:
• Gold Conjugates: Mouse monoclonal anti-MET gold colloid (0.47 pi)
Mouse monoclonal anti-THC-gold colloid (0.47 pi)
• Test LineM:
Methamphetamine-BSA conjugate(0.23 pg)
• Test Line T:
THC-BSA conjugate (1.17 pg)
• Control Line: Goat anti-mouse IgG (0.35 pg)
Other materials required for testing procedure:
• Protective gloves
• Timer
• Biohazard container
IV.
TESTING PROCEDURE
(1) Bring all kit components and specimens to reach a temperature between 15°C and 30 °C prior to testing.
(2) Remove the test device from foil pouch and place it on a flat, dry surface. Label the test device with a
patient identifier.
(3) Holding the urine dropper above the test device, dispense 3 - 4 drops of urine into the sample well (S).
(4) As the test begins to work, you will see a purple colored band move across the result window in the center
of the test device.
(5) Interpret test results at 5 minutes. Do not interpret test result after 5 minutes.
V.
RESULTS
(1) Negative result: The presence of three colored lines (M, T and C) within the result window, regardless of the
order in which the lines appear, indicates a negative result. A negative result indicates that the drug
concentrations are below the detectable levels.
(2) Methamphetamine positive result: The presence of both the THC test line (T) and the control line (C) within
the result window, regardless of which line appears first, indicates a methamphetamine-positive result. This
result indicates that the concentration of methamphetamine in the sample is at or above the detectable
level (1,000 ng/ml).
(3) THC positive result: The presence of both the MET test line (M) and the control line (C) within the result
window, regardless of which line appears first, indicates a THC-positive result. This result indicates that the
concentration of THC metabolites in the sample is at or above the detectable level (50 ng/ml).
(4) Methamphetamine and THC positive result: The presence of only the control line (C) within the result
window indicates a positive result for both methamphetamine and THC. This result indicates that the
concentrations of methamphetamine and THC metabolites are at or above their detectable levels (1,000
ng/ml and 50 ng/ml, respectively).
(5) Invalid result: If the control line (C) is not visible within the result window after performing the test, the
result is considered invalid. Instructions may not have been followed correctly or the test may have
deteriorated. It is recommended that the specimen be retested using a new test device.
VI.
TEST LIMITATIONS
• The Bioline MET/THC test is designed for use with human urine only.
• The test provides only a preliminary analytical result. A secondary analytical method must be used to
obtain a confirmatory result. Gas chromatography/mass spectrometry (GC/MS) is the preferred
confirmatory method.
• The Bioline MET/THC test is a qualitative screening assay and cannot determine the concentration of
methamphetamine or THC metabolites in the urine or the level of intoxication.
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VII.
It is possible that technical or procedural errors, as well as other interfering substances in the urine
specimen, may cause erroneous results.
Adulterants such as bleach and/or alum in urine specimens may yield suspected, the test should be
repeated with another urine specimen.
Certain medications containing opiates or opiate derivatives may produce a positive result. Foods and tea
containing poppy products may also produce a positive result.
QUALITY ASSURANCE AND CONTROL
A. PROPER KIT STORAGE AND INSPECTION
• The test kit should be stored at a temperature between 2°C and 30 °C. Do not freeze the kit or its
components.
• The test device is sensitive to both heat and humidity. Perform the test immediately after removing the
test device from foil pouch.
• Do not use the test kit beyond its expiration date. The shelf life of the kit is as indicated on the outer
package.
• Do not use the test kit if the pouch is damaged or the seal is broken.
B. USE OF INTERNAL QUALITY CONTROL
• The presence of a colored band in the control region (C) acts as the internal procedural control included
in the test. It confirms sufficient specimen volume and correct procedural technique. A clear background
is required.
• Control standards are not supplied with this kit. However, good laboratory testing practice recommends
that positive and negative controls be tested to confirm the test procedure and to verify proper test
performance; the US National Institute for Drug Abuse (NIDA) recommends that positive quality control
specimens be at or near the cut-off concentrations.
C. PRECAUTIONS
• The test devices are for in vitro diagnostic use only. Do not reuse the test device.
• Read the entire instruction for use carefully before using this test. If you don’t follow this instruction for
use (including precautions and test procedure), you may obtain false result and make foreseeable misuse.
This test works only if the instructions are followed exactly.
• Avoid cross contamination of urine specimens by using a new urine specimen container and dropper for
each urine specimen.
• Do not use the pipette by mouth, smoke, drink, eat, apply cosmetics, or handle contact lenses in areas
where specimens or kit components are being handled.
• Wear protective gloves while handling specimens and wash hands thoroughly afterwards.
• Clean up spills thoroughly using an appropriate disinfectant.
• Decontaminate and dispose of all specimens, reaction kits and potentially contaminated materials in a
biohazard container as if they were infectious waste.
• Do not mix or interchange different specimens.
• Do not eat the desiccant from the foil pouch.
• Do not mix or interchange components among different lots or those for other products.
VIII.
PERFORMANCE CHARACTERISTICS
A. EXPECTED VALUES
• This kit can detect methamphetamine and THC metabolites in human urine. A positive result indicates
that the sample probably contains methamphetamine and/or Δ9-THC-COOH at or above the
concentrations of 1,000 ng/ml and 50 ng/ml, respectively.
B. SENSITIVITY AND SPECIFICITY
• The NIDA has suggested that the screening cut-off for positive samples be 1,000ng/ml for
methamphetamine and 50ng/ml for Δ9-THC-COOH.
• The accuracy of the Bioline MET/THC test was evaluated in comparison to a commercially available test
(immunoassay) at the cut-off of 1,000 ng/ml of methamphetamine and 50 ng/ml of Δ9-THC-COOH. Bioline
MET/THC test kit is designed to have
o 100 % (95 % Cl: 94.5 - 100 %) of sensitivity and 100 % (95 % Cl: 96.9 - 100 %) of specificity for MET.
o 100 % (95 % Cl:94.6 - 100 %) of sensitivity and 100 % (95 % Cl: 96.9 - 100 %) of specificity for THC
C. REPRODUCIBILITY
• Reproducibility of the Bioline MET/THC test has been demonstrated by within-run, between-run and
batch-to-batch studies using in-house reference panels. All values were identical to reference panel
acceptance criteria.
D. ANALYTICAL SENSITIVITY
• The Bioline MET/THC can detect 1,000 ng/ml of methamphetamine and 50 ng/ml of A9-THC-COOH as its
confirmed minimum detection limits. The Bioline MET/THC test is equal to other commercial kits for the
detection of low concentrations of methamphetamine and THC metabolites.
E. ANALYTICAL SPECIFICITY
• Due to cross-reactivity resulting from the interaction of efavirenz metabolites with the THC rapid test, it
is possible that anti-retroviral therapy with efavirenz produces urine samples that screen false positive for
THC, despite the absence of THC metabolites.
• The following table lists compounds that are positively detected in urine by the Bioline MET/THC test at 5
minutes:
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IX.
The following compounds showed no cross-reactivity when tested with the Bioline MET/THC test at a
concentration of 100 pg/ml:
REFERENCE
SD Bioline METR/THC Kit insert
Abbott Diagnostic, Korea
Date issued: 2021. 03
S0FK60-02-EN.ES-A1