VITROS CHEMISTRY PRODUCTS OP REAGENT, CALIBRATOR KIT 26 AND DAT PERFORMANCE VERIFIERS I, II AND III

{0} 1 # 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY ASSAY ONLY TEMPLATE A. 510(k) Number: k062460 B. Purpose for Submission: New device C. Measurand: Opiates D. Type of Test: Qualitative and semi-quantitative enzyme immunoassay E. Applicant: Ortho-Clinical Diagnostics, Inc. F. Proprietary and Established Names: VITROS Chemistry Products OP Reagent VITROS Chemistry Products Calibrator 26 VITROS Chemistry Products FS Calibrator 1 VITROS Chemistry Products DAT Performance Verifiers I, II, III, IV and V G. Regulatory Information: 1. Regulation section: 21 CFR 862.3650 Opiate test system 21 CFR 862.3200, Clinical Toxicology Calibrator 21 CFR 862.3180, Clinical Toxicology Control 2. Classification: II (reagent, calibrator) I, reserved (control) 3. Product code: DJG, DKB and DIF {1} 4. Panel: Toxicology (91) H. Intended Use: 1. Intended use(s): See Indications for use. 2. Indication(s) for use: VITROS Chemistry Products OP Reagent: For in vitro diagnostic use only. VITROS Chemistry Products OP Reagent is used on VITROS 5,1 FS Chemistry Systems for the semi-quantitative or qualitative determination of opiates (OP) in human urine using a cutoff of either 300 ng/mL or 2000 ng/mL. Measurements obtained with the VITROS OP method are used in the diagnosis and treatment of opiate use or overdose. The VITROS Chemistry Products OP 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 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: For in vitro diagnostic use only. 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 I, II, III, IV &amp; V: For in vitro diagnostic use only. 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): For use by professional laboratory personnel. For in vitro diagnostic use only. 4. Special instrument requirements: Ortho-Clinical Diagnostics VITROS 5,1 FS Chemistry System I. Device Description: The VITROS Chemistry Products OP Reagent is a dual chambered reagent pack containing two ready-to-use liquid reagents. The reactive ingredients in Reagent 1 include sheep polyclonal antibodies reactive to morphine, Glucose 6-phosphate and 2 {2} Nicotinamide adenine nucleotide (NAD). The other ingredients in Reagent 1 include inorganic salt, organic salt, proteins, inorganic polymer, protease inhibitor, stabilizer, surfactant and preservative. The reactive ingredients in Reagent 2 include morphine labeled with glucose-6-phosphate dehydrogenase. The other ingredients in Reagent 2 include buffers, organic salt, inorganic salt, proteins, inorganic polymer, protease inhibitor, biological material, surfactant and preservatives. 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 PCP Reagent on VITROS 5,1 FS Chemistry Systems. The product labeling for Calibrator Kit 26 and Performance Verifiers contain warnings regarding the presence of human source materials and recommend the use of Universal Precautions when handling these products. ## J. Substantial Equivalence Information: 1. Predicate device name(s): Syva EMIT II Plus Opiate assay Bio-Rad Liquicheck Urine Toxicology Controls 2. Predicate 510(k) number(s): k011289 k022707 3. Comparison with predicate: | Similarities | | | | --- | --- | --- | | Item | Device | Predicate | | Intended Use | For use in the qualitative and semi-quantitative analysis of opiates in human urine. | Same | | Reagent | Liquid, ready to use | Same | | Principle | Homogeneous enzyme immunoassay | Same | | Matrix | Urine | Same | | Antibody | Sheep polyclonal | Same | {3} | Differences | | | | --- | --- | --- | | Item | Device | Predicate | | Instrumentation | VITROS 5,1 FS Chemistry Systems | Multiple automated clinical chemistry analyzers | | Calibrators | Six levels | Qualitative: two levels Semi-quantitative: 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 OP assay is a homogenous immunoassay based on the competition between morphine in the treated urine sample and morphine 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 morphine 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. ## M. Performance Characteristics (if/when applicable): ### 1. Analytical performance: a. Precision/Reproducibility: Precision was evaluated with quality control materials on the VITROS 5,1 FS Chemistry System following CLSI EP5-A. The samples were run in duplicate, twice a day for twenty-two days using two reagent lots and four instruments. The results are presented in the table below. {4} | Semi-quantitative-LO (300) | | | | | | | --- | --- | --- | --- | --- | --- | | Mean Conc. ng/mL | Within-Day SD | Within-Lab SD | Within-Lab %CV | No. Observations | No. Days | | 213 | 5.1 | 10.5 | 4.9 | 88 | 22 | | 294 | 5.4 | 12.4 | 4.2 | 88 | 22 | | 371 | 5.5 | 14.5 | 3.9 | 88 | 22 | | Semi-quantitative-HI (2000) | | | | | | | --- | --- | --- | --- | --- | --- | | Mean Conc. ng/mL | Within-Day SD | Within-Lab SD | Within-Lab %CV | No. Observations | No. Days | | 1459 | 24.1 | 65.2 | 4.5 | 88 | 22 | | 1995 | 37.3 | 88.0 | 4.4 | 80 | 20 | | 2441 | 46.4 | 112.9 | 4.6 | 88 | 22 | | 2901 | 62.7 | 139.2 | 4.8 | 88 | 22 | Qualitative imprecision was assessed using drug-spiked human urine pools with concentrations targeted at approximately $\pm 25\%$ of the 300 and $2000~\mathrm{ng/mL}$ cutoff concentrations. The concentrations of the targeted test fluids were confirmed by GC/MS. The sponsor performed two runs per day with two replicates per run for 22 days using a single lot of reagent on one analyzer. The results are presented below: | Qualitative | | | | | | --- | --- | --- | --- | --- | | Cutoff ng/mL | Test Fluid Concentration ng/mL | Test Fluid % of Cutoff | No. Observations | No. of Correct Results | | 300 | 213 | -25% of the cutoff | 88 | 88 | | 300 | 371 | +25% of the cutoff | 88 | 88 | {5} | Qualitative | | | | | | --- | --- | --- | --- | --- | | Cutoff ng/mL | Test Fluid Concentration ng/mL | Test Fluid % of Cutoff | No. Observations | No. of Correct Results | | 2000 | 1459 | -25% of the cutoff | 88 | 88 | | 2000 | 2441 | +25% of the cutoff | 88 | 88 | b. Linearity/assay reportable range: The sponsor followed CLSI EP6-A in determining the linear range of their device. Number of reagent lots: 3 Replicates of each solution: 3 OP-LO (300 ng/mL) Two urine pools were prepared with a high urine based morphine pool (4000 ng/mL) and a negative urine based pool (0 ng/mL). The two pools were mixed to give 16 admixtures of intermediate concentrations. Linearity was evaluated using three 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 concentration range tested (69 to 1009 ng/mL). The claimed reportable range of the VITROS OP-LO assay is 100 - 900 ng/mL. OP-HI (2000 ng/mL) Two urine pools were prepared with a high urine based morphine pool (5000 ng/mL) and a negative urine based pool (0 ng/mL). The two pools were mixed to give 11 admixtures of intermediate concentrations. Linearity was evaluated using three assay reagent lots and comparing the measured results against the expected results from 11 pooled samples. A linear regression was performed and the results indicated acceptable linearity across the concentration range tested (503 to 4000 ng/mL). The claimed reportable range of the VITROS OP-HI assay is 700 - 4000 ng/mL. c. Traceability, Stability, Expected values (controls, calibrators, or methods): The assigned values for the calibrators are traceable to the Cerilliant morphine standard catalogue M-109 and are verified by GC/MS. The assigned values for the controls are traceable to USP 14447002 (Morphine Monohydrate CII). They are prepared through gravimetric addition to drug free urine and verified by GC/MS. Real time and accelerated stability studies were conducted; protocols and {6} acceptance criteria were described and found to be acceptable. These studies support the manufacturer's stability claims for the following products: | Reagent | Storage | Stability* | | --- | --- | --- | | Unopened | 2-8°C | 5 months | | Opened | On board analyzer, system turned off | ≤28 days | | Opened | On board analyzer, system turned on | ≤30 minutes | | Calibrator | Storage | Stability* | | --- | --- | --- | | Unopened | ≤18°C | 8 months | | Opened | 2-8°C | 4 weeks | | Controls | Storage | Stability* | | --- | --- | --- | | Unopened | 2-8°C | 6 months | | Opened | 2-8°C | 4 weeks. | *Note: Real time studies are ongoing. d. Detection limit: The detection limit was determined according to protocol recommendations in CLSI EP-17 on three different lots of reagent and one instrument platform. The claimed lowest detectable limit for VITROS OP-LO is $97\mathrm{ng / mL}$ . The claimed lowest detectable limit for VITROS OP-HI is $338\mathrm{ng / mL}$ . e. Analytical specificity: The sponsor conducted interference studies following CLSI EP7-A2. The substances listed in the table below were determined not to interfere in the concentration tested at $300\mathrm{ng / mL}$ morphine and $2000\mathrm{ng / mL}$ morphine cutoffs, up to the concentrations shown: | Compound | Concentration Tested | | | --- | --- | --- | | | Conventional | SI | | ammonia | 570 mg/dL | 316 mmol/L | | ascorbic Acid | 500 mg/dL | 28.4 mmol/L | | Bilirubin | 26 mg/dL | 444 μmol/L | | buprenorphine | 5 mg/dL | 106 μmol/L | | Calcium | 30 mg/dL | 7.5 mmol/L | | ciprofloxacin | 10 mg/dL | 302 μmol/L | {7} | Compound | Concentration Tested | | | --- | --- | --- | | | Conventional | SI | | citric acid | 100 mg/dL | 5.2 mmol/L | | Cloxacillin | 10 mg/dL | 230 μmol/L | | creatinine | 300 mg/dL | 26.5 mmol/L | | Diethylpropione | 10 mg/dL | 487 μmol/L | | ethacrylnic acid | 10 mg/dL | 330 μmol/L | | Ethanol | 780 mg/dL | 169 mmol/L | | Glucose | 4000 mg/dL | 222 mmol/L | | Hemoglobin | 500 mg/dL | 5 g/L | | Human IgG | 200 mg/dL | 2 g/L | | Human serum albumin | 200 mg/dL | 2 g/L | | Indomethacin | 10 mg/dL | 279 μmol/L | | Iron | 0.1 mg/dL | 17.9 μmol/L | | Magnesium | 60 mg/dL | 24.7 mmol/L | | Methoxyphenamine | 10 mg/dL | 558 μmol/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 | | Potassium | 587 mg/dL | 150 mmol/L | | Propoxyphene | 10 mg/dL | 295 μmol/L | | Propanolol | 10 mg/dL | 385 μmol/L | | Pyruvate | 200 mg/dL | 22.8 mmol/L | | Ranitidine | 10 mg/dL | 318 μmol/L | | Riboflavin | 2 mg/dL | 53 μmol/L | | Tolmetin/tolectin | 10 mg/dL | 390 μmol/L | | Trihexylphenidyl | 10 mg/dL | 332 μmol/L | | Trimethobenzamide | 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 | The sponsor determined that a high specific gravity does not interfere with the assay by evaluating the primary causes of high specific gravity: high concentrations of NaCl, protein, and glucose in urine. The specificity of VITROS OP assay for morphine and structurally similar compounds was determined by generating a dose response curve for each of the compounds and determining the approximate quantity of each compound that is equivalent in assay reactivity to the $300\mathrm{ng / mL}$ and $2000\mathrm{ng / mL}$ {8} cutoffs. | Compound | Quantity equivalent to 300 ng/mL | Approx. % Cross-reactivity | Quantity equivalent to 2000 ng/mL | Approx. % Cross-reactivity | | --- | --- | --- | --- | --- | | Ethylmorphine | 165 | 181.8% | 1175 | 170.2% | | Codeine | 190 | 157.9% | 950 | 210.5% | | Hydrocodone | 240 | 125% | 1510 | 132.5% | | Dihydrocodeine | 250 | 120% | 1420 | 140.8% | | Morphine | 300 | 100% | 2000 | 100% | | Thebaine | 400 | 75% | 2900 | 69% | | Hydromorphone | 420 | 71.4% | 4100 | 48.8% | | 6-acetyl morphine | 430 | 69.8% | 3920 | 51% | | Heroin (diacetylmorphine) | 490 | 61.2% | 3850 | 51.9% | | Dihydromorphone | 535 | 56.1% | 6025 | 33.2% | | Levorphanol | 550 | 54.5% | 10800 | 18.5% | | Morphine-3-glucuronide | 700 | 42.9% | 11100 | 18% | | Oxycodone | 2175 | 13.8% | 56500 | 3.5% | | Norcodeine | 2840 | 10.6% | 50000 | 4% | | Nalorphine | 10025 | 3% | 290000 | 0.7% | | Oxymorphone | 12600 | 2.4% | >200000 | <1% | | Normorphine | 24000 | 1.3% | 458000 | 0.4% | | Meperidine | 33500 | 0.9% | 459000 | 0.4% | | Naloxone | 466000 | 0.1% | >1000000 | <0.2% | | Fluphenazine | >100,000 | <0.3% | >1000000 | >0.2% | f. Assay cut-off: There are two cutoffs for this assay: 300 ng/mL and 2000 ng/mL. The 2000 ng/mL cutoff concentration is currently recommended by the Substance Abuse and Mental Health Services Administration (SAMHSA). Characterization of how the device performs analytically around the claimed cutoff concentrations appear in the precision section, above. 2. Comparison studies: a. Method comparison to the predicate device The results obtained on the new device were compared to the reference method, Gas Chromatography/Mass Spectrometry, and a commercially marketed (predicate) device. Results are summarized below: {9} 10 # OP-LO 300 ng/mL cutoff One hundred and eighteen unaltered human urine samples were assayed on the device and the results were compared to GC/MS. The results are corrected for cross-reactivity. ## Comparison of VITROS OP-LO to GC/MS is presented in the following table. | Cutoff ng/mL | | <50% <150 ng/mL | -50% to cutoff 150-300 ng/mL | Cutoff to +50% 300-450 ng/mL | >+50% >450 ng/mL | % Agreement Negative | % Agreement Positive | % Agreement Overall | | --- | --- | --- | --- | --- | --- | --- | --- | --- | | 300 | VITROS + | 0 | 2* | 9 | 55 | 95.8 | 91.4 | 93.2 | | | VITROS - | 29 | 17 | 5* | 1* | | | | *The summary of discordant results is presented in the table below: | Discordant Results | | | | --- | --- | --- | | Cutoff ng/mL | VITROS OP-LO ng/mL | GC/MS ng/mL | | 300 | 333 | 268 Morphine | | | 303 | 277 Morphine | | | 295 | 311 Morphine | | | 277 | 331 Morphine | | | 270 | 368 Morphine | | | 282 | 376 Morphine | | | 296 | 313 Morphine 69 Hydromorphone | | | 280 | 463 Morphine | # OP-HI 2000 ng/mL cutoff One hundred and fifty three unaltered human urine samples were assayed on the device and the results were compared to GC/MS. The results are corrected for cross-reactivity. ## Comparison of VITROS OP-HI to GC/MS is presented in the following table. | Cutoff ng/mL | | <50% <1000 ng/mL | -50% to cutoff 1000-2000 ng/mL | Cutoff to +50% 2000-3000 ng/mL | >+50% >3000 ng/mL | % Agreement Negative | % Agreement Positive | % Agreement Overall | | --- | --- | --- | --- | --- | --- | --- | --- | --- | {10} | Cutoff ng/mL | | <50% <1000 ng/mL | -50% to cutoff 1000-2000 ng/mL | Cutoff to +50% 2000-3000 ng/mL | >+50% >3000 ng/mL | % Agreement Negative | % Agreement Positive | % Agreement Overall | | --- | --- | --- | --- | --- | --- | --- | --- | --- | | 2000 | VITROS + | 1* | 1* | 5 | 35 | 98.1 | 81.6 | 92.8 | | | VITROS - | 87 | 15 | 8* | 1* | | | | *The summary of the discordant results is presented in the table below: | Discordant Results | | | | --- | --- | --- | | Cutoff ng/mL | VITROS OP-HI ng/mL | GC/MS ng/mL | | 2000 | 2669 | 421 Morphine | | | 3986 | 720 Hydrocodone 971 Hydromorphone | | | 1296 | 2090 Morphine | | | 1428 | 2330 Morphine | | | 1668 | 2373 Morphine | | | 1292 | 2425 Morphine | | | 1270 | 2612 Morphine | | | 1898 | 2593 Morphine 113 Hydromorphone | | | 1797 | 2690 Morphine | | | 1605 | 2826 Morphine | | | 1653 | 3440 Morphine | The sponsor notes that the primary form of morphine in urine is morphine-3-glucuronide, a morphine conjugate. The VITROS Chemistry Product OP Assay has lower recovery with morphine conjugates than with free morphine. As part of sample preparation for GC/MS analysis, morphine conjugates are commonly hydrolyzed to morphine. This may lead to an apparent under-recovery of opiates when comparing VITROS OP Assay results to GC/MS. ## OPI-LO 300 ng/mL cutoff One hundred and twenty one human urine samples were assayed on the device and were compared to results from the predicate device. {11} Comparison of VITROS OP-LO to the Predicate Device is presented in the following table. | Cutoff ng/mL | | <50% <150 ng/mL | -50% to cutoff 150-300 ng/mL | Cutoff to +50% 300-450 ng/mL | >+50% >450 ng/mL | % Agreement Negative | % Agreement Positive | % Agreement Overall | | --- | --- | --- | --- | --- | --- | --- | --- | --- | | 300 | VITROS + | 0 | 0 | 5 | 63 | 100 | 93.2 | 95.9 | | | VITROS - | 27 | 21 | 5* | 0 | | | | *The summary of discordant results is presented in the table below. | Discordant Results | | | | --- | --- | --- | | Cutoff ng/mL | VITROS OP-LO ng/mL | Predicate Assay ng/mL | | 300 | 251 | 321 | | | 261 | 304 | | | 284 | 333 | | | 295 | 399 | | | 296 | 414 | OPI-HI: 2000 ng/mL cutoff One hundred and fifty five human urine samples were assayed on the device and were compared to results from the predicate device. Comparison of VITROS OP-HI to the Predicate Device are presented in the following table. | Cutoff ng/mL | | <50% <1000 ng/mL | -50% to cutoff 1000-2000 ng/mL | Cutoff to +50% 2000-3000 ng/mL | >+50% >3000 ng/mL | % Agreement Negative | % Agreement Positive | % Agreement Overall | | --- | --- | --- | --- | --- | --- | --- | --- | --- | | 2000 | VITROS | 0 | 1* | 8 | 34 | 99.1 | 91.3 | 96.8 | | | VITROS | 77 | 31 | 4* | 0 | | | | {12} The summary of the discordant results is presented in the table below. | Discordant Results | | | | --- | --- | --- | | Cutoff ng/mL | VITROS OP-HI ng/mL | Predicate ng/mL | | 2000 | 1653 | 2107 | | | 1797 | 2135 | | | 1898 | 2472 | | | 1993 | 2098 | | | 2005 | 1966 | b. Matrix comparison: Not applicable 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.