HANDLE ALL URINE SPECIMENS AS IF THEY ARE POTENTIALLY INFECTIOUS. Catalog Number: 311UR-0025 25 mL Kit Catalog Number: 311UR-0100 100 mL Kit ASSAY PROCEDURE Catalog Number: 311UR-0500 500 mL Kit Analyzers capable of maintaining a constant temperature, pipetting samples and Catalog Number: 311UR-0060W 60 mL Kit reagents, mixing reagents, timing the reaction accurately and measuring enzymatic Intended Use: rates at 340 nm can be used to perform the assay. The assay has been validated for The Immunalysis Methamphetamine Urine Enzyme Immunoassay is a homogeneous use with the Olympus AU400e, the following Beckman Coulter analyzers: AU480, enzyme immunoassay with a dual cutoff of 500ng/mL and 1000ng/mL. The assay is AU680, AU2700, AU5400 and AU5800. intended for use in laboratories for the qualitative and semi-quantitative analysis of Methamphetamine in human urine with automated clinical chemistry analyzers. This assay Refer to the analyzer-specific Application Sheet which contains parameters for use. is calibrated against Methamphetamine. This in-vitro diagnostic device is for prescription QUALITY CONTROL AND CALIBRATION use only. Good laboratory practice suggests the use of control specimens to ensure assay The semi-quantitative mode is for purposes of enabling laboratories to determine an performance. Control results must fall within established ranges determined by your appropriate dilution of the specimen for confirmation by a confirmatory method such as laboratory. QC materials should be used in accordance with local, state and/or federal Gas Chromatography/ Mass Spectrometry (GC-MS) or permitting laboratories to establish regulations or accreditation requirements. quality control procedures. For a qualitative analysis use either the 500ng/mL or 1000ng/mL calibrator as a cutoff The Immunalysis Methamphetamine Urine Enzyme Immunoassay Kit provides only level to distinguish “positive” and “negative” specimens. For semi-quantitative analysis a preliminary analytical test result. A more specific alternate chemical method must use all calibrators. be used in order to obtain a confirmed analytical result. GC-MS or Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is the preferred RESULTS AND EXPECTED VALUES confirmatory method. Clinical consideration and professional judgment should be Qualitative Results applied to any drug of abuse test result, particularly when preliminary positive A specimen that exhibits a change in absorbance ( ∆A) value, equal to or greater than results are used. the value obtained with the cutoff calibrator is considered positive. A sample that EXPLANATION OF THE TEST exhibits a change in absorbance ( ∆A) value, lower than the value obtained with the [1] Methamphetamine (USAN) also known as metamfetamine (INN), meth, ice, speed, crystal, cutoff calibrator is considered negative. glass, tik, N-methylamphetamine, methylamphetamine, and desoxyephedrine, is a Semi-quantitative Results psychostimulant of the phenethylamine and amphetamine class of psychoactive drugs. A rough estimate of drug concentration in the specimens can be obtained by running a Methamphetamine increases alertness, concentration, energy, and in high doses, can induce standard curve with all calibrators and calculating samples of the standard curve. [2][3] euphoria, enhance self-esteem and increase libido. Methamphetamine has a high potential for abuse and addiction, activating the psychological reward system by triggering a cascading release LIMITATIONS of dopamine in the brain. Methamphetamine is metabolized in the liver with the main metabolites • The assay is designed for use with human urine only. being amphetamine (active) and 4-hydroxymethamphetamine (pholedrine). It is excreted by the • Boric Acid at 1% w/v may cause false negative results. Boric Acid is not kidneys, with the rate of excretion into the urine heavily influenced by urinary pH. Between 30- recommended as a preservative for urine. 54% of an oral dose is excreted in urine as unchanged methamphetamine and 10-23% as • Other substances and factors not listed may interfere with the test and cause false unchanged amphetamine. Following an intravenous dose, 45% is excreted as unchanged parent [4] results. drug and 7% amphetamine. The half-life of methamphetamine is variable with a mean value of • [5] Interpretation of results must take into account that urine concentrations can vary between 9 and 12 hours. extensively with fluid intake and other biological variables. Immunoassays that PRINCIPLES OF THE PROCEDURE produce a single result in the presence of a drug and its metabolites cannot fully This assay uses Methamphetamine recombinant antibody. The assay is based on the quantitate the concentration of individual components. competition of Methamphetamine labeled enzyme glucose-6-phosphate dehydrogenase (G6PDH) and the free drug in the urine sample for the fixed amount of antibody binding SPECIFIC PERFORMANCE CHARACTERISTICS sites. In the absence of the free drug in the sample, the antibody binds the drug enzyme Performance results obtained on the Olympus AU400e analyzer are shown below. conjugate and enzyme activity is inhibited. This creates a dose response relationship Additionally, the following analyzers have been validated for use with the between drug concentration in the urine and enzyme activity. The enzyme G6PDH activity Methamphetamine Urine Enzyme Immunoassay. is determined at 340 nm spectrophotometrically by the conversion of NAD to NADH. Manufacturer Analyzer REAGENTS Beckman Coulter AU480 Beckman Coulter AU680 Immunalysis Methamphetamine Urine Enzyme Immunoassay Kit Beckman Coulter AU2700 Beckman Coulter AU5400 Antibody/Substrate Reagent (RA) - This contains recombinant antibodies to Beckman Coulter AU5800 Methamphetamine, glucose-6-phosphate (G6P) and nicotinamide adenine dinucleotide Refer to the analyzer-specific Application Sheet for information on analyzer settings (NAD) in Tris buffer with Sodium Azide as a preservative. and parameters. Enzyme Conjugate Reagent (RE) - This contains glucose-6-phosphate dehydrogenase Semi-Quantitative Precision was determined by testing calibrators and controls for 20 (G6PDH) labeled with Methamphetamine in Tris buffer with Sodium Azide as a days, 2 runs per day in replicates of 2 (N=80). preservative. Table 1: Semi-Quantitative Precision Additional Materials Required (sold separately): 500ng/mL Cutoff Catalog No.: Concentration (ng/mL) Result Total Result 10000 -15 or Immunalysis Multi -Drug Calibrators Level 1 Calibrator containing 150ng/mL 125ng/mL 10000-25 of Benzoylecgonine, 500ng/mL of Methamphetamine, 100ng/mL of NEG 80 Negative Morphine, 12.5ng/mL of PCP and 100ng/mL of Oxazepam 250ng/mL NEG 80 Negative 10001 -15 or Immunalysis Multi -Drug Calibrators Level 2 Calibrator containing 300ng/mL 375ng/mL (Control LOW) NEG 80 Negative 10001-25 of Benzoylecgonine, 1000ng/mL of Methamphetamine, 300ng/mL of 500ng/mL Calibrator N/A 35 Negative/45 Positive Morphine, 25g/mL of PCP and 200ng/mL of Oxazepam 625ng/mL (Control HIGH) POS 80 Positive 10002 -15 or Immunalysis Multi -Drug Calibrators Level 3 Calib rator containing 500ng/mL 750ng/mL POS 80 Positive 10002-25 of Benzoylecgonine, 1500ng/mL of Methamphetamine, 500ng/mL of 875ng/mL POS 80 Positive Morphine, 50ng/mL of PCP and 500ng/mL of Oxazepam 1000ng/mL POS 80 Positive 10003 -15 or Immunalysis Multi -Drug Calibrators Level 4 Calibrator containing 1000ng/mL 1000ng/mL Cutoff 10003-25 of Benzoylecgonine, 2000ng/mL of Methamphetamine, 1000ng/mL of Concentration (ng/mL) Result Total Result Morphine, 100ng/mL of PCP and 1000ng/mL of Oxazepam 3000 -15 o r 250ng/mL NEG 80 Negative Immunalysis Multi-Drug Control Set 1 containing LOW and HIGH Control 1 3000-25 500ng/mL NEG 80 Negative 3001 -15 or Immunalysis Multi -Drug Control Set 2 containing LOW and HIGH Control 2 750ng/mL (Control LOW) NEG 80 Negative 3001-25 1000ng/mL Calibrator N/A 37 Negative/43 Positive 1250ng/mL (Control HIGH) POS 80 Positive WARNING 1500ng/mL POS 80 Positive 1) H303 The reagents may be harmful if swallowed. 1750ng/mL POS 80 Positive 2) H290 Reagents contain Sodium Azide as a preservative. When disposing of these 2000ng/mL POS 80 Positive reagents flush with large volumes of water to prevent azide build-up which could react with See Application Sheet for calibration frequency recommendation. copper and lead plumbing to form potentially explosive metal azides. 3) Do not use reagents beyond their expiration dates. Qualitative Precision was determined by testing calibrators and controls for 20 Preparation and Storage days, 2 runs per day in replicates of 2 (N=80). The Immunalysis Methamphetamine Urine Enzyme Immunoassay Kit reagents are Table 2: Qualitative Precision provided ready to use and may be used directly from the refrigerator. 500ng/mL Cutoff Reagent bottles should remain closed when not in use with screw caps tightly Concentration (ng/mL) Result Total Result closed. The expiration date of the kit is stated on the label. The expiration date reflects the 125ng/mL NEG 80 Negative stability claim of 18 months stored at 2 – 8°C. The reagents are stable opened when 250ng/mL NEG 80 Negative stored on-board the instrument for 28 days at 2 – 8°C 375ng/mL (Control LOW) NEG 80 Negative SPECIMEN COLLECTION - Storage and Handling Instructions 500ng/mL Calibrator Urine samples should be collected in industry standard urine collection containers. Ensure N/A 41 Negative/39 Positive that the samples are free of gross debris. Highly turbid specimens should be centrifuged 625ng/mL (Control HIGH) POS 80