VITROS CHEMISTRY PRODUCTS METD REAGENT, CALIBRATOR KIT 26, DAT PERFORMANCE, VERIFIERS I, II, III, IV & V

{0} # 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY ASSAY ONLY TEMPLATE A. 510(k) Number: k062637 B. Purpose for Submission: New product C. Measurand: Methadone D. Type of Test: Semi-quantitative and qualitative homogeneous enzyme immunoassay E. Applicant: Ortho-Clinical Diagnostic F. Proprietary and Established Names: VITROS Chemistry Products METD Reagent VITROS Chemistry Products Calibrator Kit 26 VITROS Chemistry Products DAT Performance Verifiers I. II, III, IV, and V G. Regulatory Information: 1. Regulation section: 21 CFR §862.3620, Methadone Test System 21 CFR §862.3200, Clinical Toxicology Calibrator 21 CFR §862.3280, Clinical Toxicology Control Material 2. Classification: Class II (Reagent, Calibrator) Class I, Reserved (Control) 3. Product code: DJR; DKB; DIF 4. Panel: Toxicology (91) H. Intended Use: 1. Intended use(s): See Indications for Use below. 2. Indication(s) for use: VITROS Chemistry Products METD Reagent is used on VITROS 5,1 FS Chemistry Systems for the semi-quantitative or qualitative determination of methadone (METD) in human urine using a cutoff of either 150 ng/mL or 300 ng/mL. Measurements obtained with the VITROS METD method are used in the diagnosis and treatment of methadone use or overdose. {1} The VITROS Chemistry Products METD assay is intended for use by professional laboratory personnel. It provides only a preliminary test result. A more specific alternative chemical method must be used to confirm a result obtained with this assay. Gas chromatography/mass spectrometry (GC/MS) is the preferred confirmatory method. Clinical consideration and professional judgment should be applied to any drug-of-abuse test result, particularly when evaluating a preliminary positive result. VITROS Chemistry Products Calibrator Kit 26 is used to calibrate VITROS 5.1 FS Chemistry Systems for the qualitative or semi-quantitative measurement of drugs of abuse. VITROS Chemistry Products DAT Performance Verifiers are assayed controls used to monitor performance of urine drugs of abuse screening assays on VITROS 5,1 FS Chemistry Systems. 3. Special conditions for use statement(s): This device is for use by professional laboratory personnel. For in vitro diagnostic use only. The VITROS Chemistry Products METD assay is intended for use by professional laboratory personnel. It provides only a preliminary test result. A more specific alternative chemical method must be used to confirm a result obtained with this assay. Gas chromatography/mass spectrometry (GC/MS) is the preferred confirmatory method. Clinical consideration and professional judgment should be applied to any drug-of-abuse test result, particularly when evaluating a preliminary positive result. 4. Special instrument requirements: Ortho-Clinical Diagnostics VITROS 5,1 FS Chemistry System I. Device Description: The VITROS METD Reagent is a dual chambered package containing ready-to-use liquid reagents that are used to detect methadone in urine. Sample, calibrators, and controls are automatically treated with surfactant (DAT Diluent 2) prior to addition of reagents. Treated sample is added to Reagent 1 containing antibodies reactive to methadone, glucose-6-phosphate and nicotinamide adenine dinucleotide $(\mathrm{NAD}^{+})$ , followed by Reagent 2 containing methadone labeled with the enzyme glucose-6-phosphate dehydrogenase (G6P-DH). VITROS Chemistry Products Calibrator Kit 26 is prepared from human urine to which drugs of abuse, metabolites of drugs of abuse, organic salts, surfactants and preservative have been added. VITROS DAT Performance Verifiers I, II, III, IV &amp; V are prepared from a human urine pool to which analytes, surfactant and preservative have been added. These are assayed controls used to monitor performance of the VITROS METD Reagent on VITROS 5,1 FS Chemistry Systems. The product labeling for the Calibrator Kit 26 and Performance Verifiers contain warnings regarding the presence of human sourced materials and recommend the use {2} of Universal Precautions when handling these products. ## J. Substantial Equivalence Information: 1. Predicate device name(s): EMIT II Plus Methadone Assay Liquicheck Urine Toxicology Controls 2. Predicate 510(k) number(s): k010962; k022707 3. Comparison with predicate: | Similarities | | | | --- | --- | --- | | Item | Device | Predicate | | Intended Use | For use in the qualitative and semi-quantitative analysis of methadone in human urine. | Same | | Reagent | Liquid, ready to use | Same | | Principle | Homogeneous enzyme immunoassay | Same | | Matrix | Urine | Same | | Antibody | Sheep polyclonal | Same | | Differences | | | | --- | --- | --- | | Item | Device | Predicate | | Instrumentation | VITROS 5,1 FS Chemistry Systems | Multiple automated clinical chemistry analyzers | | Calibrators | Six levels | Four levels | | Controls | Five levels | Two levels | ## K. Standard/Guidance Document Referenced (if applicable): CSLI EP9-A2: Method Comparison and Bias Estimation Using Patient Samples CLSI EP5-A: Evaluation of Precision Performance of Clinical Chemistry Devices CLSI EP6-A: Evaluation of the Linearity of Quantitative Measurement Procedures, A Statistical Approach CLSI EP7-P: Interference Testing in Clinical Chemistry CLSI EP17-A: Protocols for Demonstration, Verification and Evaluation of Limits of Detection and Quantitation CLSI EP12-A: User Protocols for Evaluation of Qualitative Test Performance ## L. Test Principle: The VITROS METD Reagent is a dual chambered package containing ready-to-use liquid reagents that are used to detect methadone in urine. Sample, calibrators, and controls are automatically treated with surfactant (DAT Diluent 2) prior to addition of reagents. Treated sample is added to Reagent 1 containing antibodies reactive to methadone, glucose-6-phosphate and nicotinamide adenine dinucleotide $(\mathrm{NAD}^{+})$, {3} followed by Reagent 2 containing methadone labeled with the enzyme glucose-6-phosphate dehydrogenase (G6P-DH). The assay is based on competition between methadone in the treated urine sample and the methadone labeled with the enzyme glucose-6-phosphate dehydrogenase (G6P-DH) for antibody binding sites. Enzyme activity decreases upon binding to the antibody, therefore the concentration of methadone in the urine sample is directly proportional to measured enzyme activity. Active enzyme converts oxidized nicotinamide adenine dinucleotide $(\mathrm{NAD}^{+})$ to NADH, resulting in an absorbance change that is measured spectrophotometrically at $340~\mathrm{nm}$ . # M. Performance Characteristics (if/when applicable): # 1. Analytical performance: # a. Precision/Reproducibility: Precision was evaluated with human urine-based quality control materials on the VITROS 5,1 FS Chemistry System following CLSI Protocol EP5 and CLSI protocol EP12. Semi-Quantitative | System | Conventional Units (ng/mL) and SI Units (μg/L) | | | Within Lab CV%** | No. Observ. | No. Days | | --- | --- | --- | --- | --- | --- | --- | | | Mean Conc. | Within Day SD* | Within Lab SD** | | | | | VITROS 5,1 FS | 108 | 5.1 | 6.2 | 5.7% | 88 | 22 | | | 149 | 5.7 | 6.8 | 4.6% | 87 | 22 | | | 190 | 5.2 | 6.5 | 3.4% | 88 | 22 | | | 233 | 4.8 | 6.6 | 2.8% | 88 | 22 | | | 312 | 4.0 | 4.5 | 1.4% | 88 | 22 | | | 377 | 4.0 | 5.6 | 1.5% | 88 | 22 | | | 624 | 17.3 | 22.8 | 3.7% | 88 | 22 | * Within Day imprecision was determined using two runs per day with two replications per run. ** Within Lab imprecision was determined using a single lot of reagents with one analyzer and five calibrations. Qualitative imprecision was assessed using test fluids targeted at $\pm 25\%$ of each cutoff. The imprecision was determined as the confidence level of obtaining a correct result with known positive or negative fluids. Qualitative* | System | Cutoff Level (ng/mL & g/L) | Test Fluid at ±25% Cutoff | Number of Observations | Number of Correct Interpretations | Confidence Level | | --- | --- | --- | --- | --- | --- | | VITROS 5,1 FS | 150 | -25% | 88 | 88 | >95% negative reading | | | 150 | +25% | 88 | 88 | >95% positive reading | | | 300 | -25% | 88 | 88 | >95% negative reading | {4} Qualitative* | System | Cutoff Level (ng/mL & μg/L) | Test Fluid at ±25% Cutoff | Number of Observations | Number of Correct Interpretations | Confidence Level | | --- | --- | --- | --- | --- | --- | | | 300 | +25% | 88 | 88 | >95% positive reading | * Determined using two runs per day with two replicates per run for 22 days, using a single lot of reagents with one analyzer and five calibrations b. Linearity/assay reportable range: The sponsor followed CLSI EP6-A in determining the linear range of their device. Two urine pools were prepared with methadone concentrations at the low (0 ng/mL) and high (1000 ng/mL) end of the calibration range. The two pools were mixed to give 14 admixtures of intermediate concentrations. Linearity was evaluated using two assay reagent lots and comparing the measured results against the expected results from 16 pooled samples. A linear regression was performed and the results indicated acceptable linearity across the methadone concentration range tested (12 to 736 ng/mL). The reportable range of the VITROS METD assay is 40-735 ng/mL. Recovery of Semi-Quantitative Results Eleven admixtures spanning the reportable range (40 to 735 ng/mL) were prepared from two human urine-based pools. Methadone values for the admixtures were calculated based on gravimetric addition of methadone with GC/MS verification of the high and low pools. Percent recovery was calculated using the concentration obtained by the VITROS Chemistry Products METD Assay versus the nominal methadone value. Recovery of Methadone | Nominal Methadone Concentration (ng/mL) | VITROS METD Assay (ng/mL) | % Recovery | | --- | --- | --- | | 56 | 53 | 94.8 | | 75 | 74 | 99.3 | | 100 | 98 | 98.4 | | 150 | 150 | 100.0 | | 200 | 204 | 102.2 | | 250 | 255 | 101.9 | | 300 | 297 | 99.0 | | 375 | 370 | 98.8 | | 500 | 489 | 97.8 | | 625 | 632 | 101.1 | | 700 | 693 | 99.1 | {5} c. Traceability, Stability, Expected values (controls, calibrators, or methods): The assigned values for the calibrators and controls are traceable to the Cerilliant methadone standard catalogue M-110 and are verified by GC/MS. Real time and accelerated stability studies were conducted; protocols and acceptance criteria were described and found to be acceptable. These studies support the manufacturer's stability claims. Real time studies are ongoing d. Detection limit: The detection limit was determined according to protocol recommendations in CLSI EP-17. The claimed lower limit for VITROS METD assay is $40\mathrm{ng / mL}$ e. Analytical specificity: The specificity of the VITROS Chemistry Products METD assay for various methadone metabolites and structurally similar compounds was estimated by generating a dose-response curve for each of the compounds listed below. The quantity $(\mathrm{ng / mL})$ of a compound necessary to produce a value equivalent to the methadone quantity $(\mathrm{ng / mL})$ at each cutoff value is listed below. If a sample contains more than one compound detected by the assay, lower quantities of cross-reactant than those listed below may produce a value approximately equivalent to or greater than that of the cutoff value due to their combined effects. Substances that Cross-react with METD | Compound | Quantity (ng/mL) equivalent to 150 ng/mL of methadone | % cross-reactivity * | Quantity (ng/mL) equivalent to 300 ng/mL of methadone | % cross-reactivity * | | --- | --- | --- | --- | --- | | L-A-methadol | 302 | 49.67% | 2625 | 11.43% | | LAMM | 385 | 38.96% | 5400 | 5.56% | | thioridazine | 54,000 | 0.28% | >300,000 | <0.1% | | EDDP | 171,000 | 0.09% | >300,000 | <0.1% | | sertraline | 300,000 | 0.05% | >1,000,000 | <0.03% | | EMDP | >300,000 | <0.05% | >300,000 | <0.1% | * The VITROS METD Assay cutoff value (ng/mL) divided by the amount of cross-reactant (ng/mL) that produces a value equivalent to the cutoff value, multiplied by 100. The substances listed in the table, at the concentrations shown, were tested according to CLSI Protocol EP7 and found not to interfere, bias $&lt; 28 \mathrm{ng} / \mathrm{mL}$ at $150 \mathrm{ng} / \mathrm{mL}$ methadone and bias $&lt; 57 \mathrm{ng} / \mathrm{mL}$ at $300 \mathrm{ng} / \mathrm{mL}$ methadone. Substances that Do Not Interfere with METD | Compound | Concentration | | | --- | --- | --- | | ammonia | 570 mg/dL | 316 mmol/L | | ascorbic acid | 500 mg/dL | 28.4 mmol/L | | bilirubin | 26 mg/dL | 444 μmol/L | | calcium | 30 mg/dL | 7.5 mmol/L | {6} Substances that Do Not Interfere with METD | Compound | Concentration | | | --- | --- | --- | | ciprofloxacin | 10 mg/dL | 302 μmol/L | | citric acid | 100 mg/dL | 5.2 mmol/L | | cloxacillin | 10 mg/dL | 230 μmol/L | | creatinine | 300 mg/dL | 26.5 mmol/L | | diethylproprione | 10 mg/dL | 487 μmol/L | | ethacrynic acid | 10 mg/dL | 330μmol/L | | ethanol | 780 mg/dL | 169 mmol/L | | glucose | 4000 mg/dL | 222 mmol/L | | hemoglobin | 500 mg/dL | 5.00 g/L | | human igg | 200 mg/dL | 2.00 g/L | | human serum albumin | 200 mg/dL | 2.00 g/L | | indomethacin | 10 mg/dL | 279 μmol/L | | iron | 0.1 mg/dL | 17.9 μmol/L | | potassium | 587 mg/dL | 150 mmol/L | | magnesium | 60 mg/dL | 24.7 mmol/L | | metronidazole | 10 mg/dL | 584 μmol/L | | nylidrine | 10 mg/dL | 334 μmol/L | | oxalic acid | 300 mg/dL | 23.8 mmol/L | | ph = 4 | | | | ph= 9 | | | | phenylbutazone | 10 mg/dL | 324 μmol/L | | phosphate | 950 mg/dL | 100 mmol/L | | pyruvate | 200 mg/dL | 22.8 mmol/L | | ranitidine hcl | 10 mg/dL | 318μmol/L | | riboflavin | 2 mg/dL | 53 μmol/L | | theophylline | 10 mg/dL | 555 μmol/L | | tolmetin/tolectin | 10 mg/dL | 390 μmol/L | | trimethobenzamide hcl | 10 mg/dL | 257 μmol/L | | tyramine | 10 mg/dL | 576 μmol/L | | urea | 3000 mg/dL | 499.5 mmol/L | | uric acid | 120 mg/dL | 7.14 mmol/L | f. Assay cut-off: The stated cutoff of this assay is either 150ng/mL or 300 ng/mL. 2. Comparison studies: a. Method comparison with predicate device: A total of 140 human urine samples were assayed using the VITROS Chemistry Products METD Reagent and a commercially available immunoassay method. Percent agreement was evaluated at assay cutoffs of 150 and 300 ng/mL. To challenge performance at the 150 ng/mL cutoff value, 43 of the 140 samples tested had concentrations within +/- 50% of the cutoff value. To challenge performance at the 300 ng/mL cutoff value, 56 of the 140 samples tested had concentrations within +/- 50% of the cutoff value. {7} Predicate Device Comparison - METD | Cutoff Value (ng/mL) | | Commercial Negative | Commercial Positive | % Agreement | | | | --- | --- | --- | --- | --- | --- | --- | | | | | | % Agreement Negative | % Agreement Positive | % Agreement Overall | | 150 | VITROS Positive | 1* | 91 | 98.0 | 100 | 99.3 | | | VITROS Negative | 48 | 0 | | | | | 300 | VITROS Positive | 1* | 48 | 98.9 | 100 | 99.3 | | | VITROS Negative | 91 | 0 | | | | *See Commercial Method Summary of Discordant Results below. Summary of Discordant Results: Commercial Method | Cutoff Value (ng/mL) | VITROS METD Assay (ng/mL) | Commercial Method (ng/mL) | | --- | --- | --- | | 150 | 199 | 149 | | 300 | 310 | 291 | A total of 139 human urine samples were assayed using the VITROS Chemistry Products METD Reagent and a GC/MS reference method for methadone. Percent agreement was evaluated at assay cutoff values of 150 and 300 ng/mL. To challenge performance at the 150 ng/mL cutoff value, 59 of the 139 samples tested had concentrations within +/- 50% of the cutoff value. To challenge performance at the 300 ng/mL cutoff value, 42 of the 139 samples tested had concentrations within +/- 50% of the cutoff value. GC/MS Reference Method Comparison for METD | | | GC/MS Reference Method | | | | % Agreement | | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | | Cutoff Value (ng/mL) | | Low Negative | Near Cutoff Negative | Near Cutoff Positive | High Positive | % Agreement Negative | % Agreement Positive | % Agreement Overall | | 150 | | (<-50%) <75 ng/mL | (-50% to cutoff) 75-150 ng/mL | (cutoff to +50%) 150-225 ng/mL | (>+50%) >225 ng/mL | 64.0 | 100 | 80.6 | | | VITROS Positive | 1* | 26* | 18 | 46 | | | | | | VITROS Negative | 33 | 15 | 0 | 0 | | | | {8} GC/MS Reference Method Comparison for METD | | | GC/MS Reference Method | | | | % Agreement | | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | | Cutoff Value (ng/mL) | | Low Negative | Near Cutoff Negative | Near Cutoff Positive | High Positive | % Agreement Negative | % Agreement Positive | % Agreement Overall | | 300 | | (<-50%) <150 ng/mL | (-50% to cutoff) 150-300 ng/mL | (cutoff to +50%) 300-450 ng/mL | (>+50%) >450 ng/mL | 87.5 | 100 | 90.6 | | | VITROS Positive | 1* | 12* | 13 | 22 | | | | | | VITROS Negative | 74 | 17 | 0 | 0 | | | | *See GC/MS Summary of Discordant Results below | Summary of Discordant Results: GC/MS | | | | | --- | --- | --- | --- | | Cutoff Value (ng/mL) | VITROS METD (ng/mL) | GC/MS (ng/mL) | Major Drug Present by GC/MS | | 150 | 151 | 136 | Methadone | | | 156 | 128 | | | | 160 | 141 | | | | 162 | 125 | | | | 166 | 83 | | | | 170 | 115 | | | | 171 | 132 | | | | 178 | 115 | | | | 181 | 146 | | | | 181 | 149 | | | | 182 | 108 | | | | 199 | 124 | | | | 199 | <RR | | | | 211 | 125 | | | | 228 | 130 | | | | 231 | 137 | | | | 231 | 143 | | | | 238 | 119 | | | | 246 | 82 | | | | 248 | 127 | | | | 249 | 88 | | | | 261 | 139 | | | | 265 | 102 | | | | 271 | 119 | | | | 271 | 122 | | | | 277 | 136 | | | 310 | 112 | | | | 300 | 310 | 112 | Methadone | | | 310 | 284 | | | | 314 | 269 | | | | 316 | 223 | | {9} | Summary of Discordant Results: GC/MS | | | | | --- | --- | --- | --- | | Cutoff Value (ng/mL) | VITROS METD (ng/mL) | GC/MS (ng/mL) | Major Drug Present by GC/MS | | | 320 | 219 | | | | 322 | 190 | | | | 323 | 290 | | | | 325 | 262 | | | | 327 | 253 | | | | 331 | 297 | | | | 332 | 297 | | | | 354 | 152 | | | | 586 | 292 | | b. Matrix comparison: Not applicable; this device is for use with urine only. 3. Clinical studies: a. Clinical Sensitivity: Not applicable. b. Clinical specificity: Not applicable. c. Other clinical supportive data (when a. and b. are not applicable): Not applicable 4. Clinical cut-off: Not applicable. 5. Expected values/Reference range: Not applicable. N. Proposed Labeling: The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10. O. Conclusion: The submitted information in this premarket notification is complete and supports a substantial equivalence decision.