(91 days)
Not Found
No
The device description and performance studies detail a standard enzyme immunoassay based on spectrophotometric measurement and chemical reactions, with no mention of AI or ML algorithms for data analysis or interpretation.
No
This device is an in vitro diagnostic (IVD) immunoassay intended for the qualitative and/or semi-quantitative determination of cocaine metabolite in human urine, not for treating or preventing a disease or condition.
Yes
This device is intended for the qualitative and/or semi-quantitative determination of benzoylecgonine (Cocaine Metabolite) in human urine, which provides a preliminary analytical test result used for clinical and professional judgment, indicating its use in diagnosing or aiding in the diagnosis of cocaine metabolite presence.
No
The device description clearly outlines the use of physical reagents (Reagent A and Reagent E) and a spectrophotometric method for determining enzyme activity, indicating it is a hardware-based in vitro diagnostic device, not software-only.
Yes, this device is an IVD (In Vitro Diagnostic).
Here's why:
- Intended Use: The "Intended Use / Indications for Use" section explicitly states that the assay is "intended for the qualitative and/or semi-quantitative determination of benzoylecgonine (Cocaine Metabolite) in human urine". This indicates that the device is used to test a sample taken from the human body (urine) to provide information about a physiological state (presence of cocaine metabolite).
- "For In Vitro Diagnostic Use Only": The "Intended Use / Indications for Use" section concludes with the clear statement "For In Vitro Diagnostic Use Only." This is a standard phrase used to designate a device as an IVD.
- Device Description: The description details a laboratory-based assay that uses reagents to analyze a urine sample. This aligns with the nature of in vitro diagnostics, which are performed outside of the living body.
- Performance Studies: The description of performance studies, including method comparison with LC-MS/MS and analysis of patient samples, further supports its use as a diagnostic tool for analyzing biological specimens.
N/A
Intended Use / Indications for Use
The Alinity c Cocaine assay is a homogeneous enzyme immunoassay intended for the qualitative and/or semi-quantitative determination of benzoylecgonine (Cocaine Metabolite) in human urine at a cutoff concentration of either 150 ng/mL or 300 ng/mL on the Alinity c analyzer.
The semiquantitative application is for the purpose of enabling laboratories to determine an appropriate dilution of the specimen for confirmation by a confirmatory method such as Liquid Chromatography/Tandem Mass Spectrometry (LC-MS/MS) or permitting laboratories to establish quality control procedures.
The assay provides only a preliminary analytical test result. A more specific alternative chemical method must be used to obtain a confirmed analytical result. Gas Chromatography/Mass Spectrometry (GC/MS) or Liquid Chromatography/ Tandem Mass Spectrometry (LC-MS/MS) are the preferred confirmatory method. Tests for cocaine metabolite cannot distinguish between abused drugs and certain prescribed medications.
Clinical and professional judgment should be applied to any drug of abuse test result, particularly when preliminary results are used. For In Vitro Diagnostic Use Only.
Product codes
DIO
Device Description
The DRI Cocaine Metabolite Assay is a homogeneous enzyme immunoassay using ready-to-use liquid reagents. The assay uses a specific antibody, which can detect benzoylecgonine in urine. The assay is based on the competition of an enzyme glucose-6-phosphate dehydrogenase (G6PDH) labeled drug and the drug from the urine samplefor a fixed amount of specific antibody binding sites. In the presence of free drug fromthe sample, the free drug occupies the antibody binding sites, allowing the drug-labeled G6PDH to interact with the substrate, resulting in enzyme activity. In the absence of drug from the sample, the specific antibody binds to the drug labeled with G6PDH and the enzyme activity is inhibited. This phenomenon creates a direct relationship between the drug concentration in the urine and the enzyme activity. The enzyme G6PDH activity is determined spectrophotometrically at 340 nm by measuring its ability to convert nicotinamide adenine dinucleotide (NAD) to NADH.
The assay consists of reagents (A and E).
Reagent A: Contains mouse monoclonal anti-benzoylecgonine antibody, glucose-6- phosphate (G6P), and nicotinamide adenine dinucleotide (NAD) in Tris buffer with sodium azide as preservative.
Reagent E: Contains benzoylecgonine analog labeled with glucose-6-phosphate dehydrogenase (G6PDH) in HEPES buffer with sodium azide as preservative.
Mentions image processing
Not Found
Mentions AI, DNN, or ML
Not Found
Input Imaging Modality
Not Found
Anatomical Site
Not Found
Indicated Patient Age Range
Not Found
Intended User / Care Setting
Trained professionals
Description of the training set, sample size, data source, and annotation protocol
Not Found
Description of the test set, sample size, data source, and annotation protocol
Test Set: Samples were prepared by spiking Benzoylecgonine (Cocaine Metabolite) into drug free urine at the cutoff, 25%, 50%, 75% and 100% above and below the cutoff.
Sample Size: n=120 for precision studies. At least one hundred patient samples were analyzed for method comparison/accuracy studies. For spike recovery studies, at least 21 replicates were used. Linearity study used 9 intermediate levels derived from 1000 ng/mL Benzoylecgonine spiked urine.
Data Source: Drug free urine and patient samples.
Annotation Protocol: Not Found
Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)
- Precision Study: Samples were tested in both qualitative and semi-quantitative modes. Qualitative and semi-quantitative results for 150 ng/mL and 300 ng/mL cutoffs were summarized, showing the number of determinants and immunoassay results (Negative/Positive) for various spiked concentrations. For the 150 ng/mL cutoff, at 150 ng/mL, 76/44 (qualitative) and 73/47 (semi-quantitative) were observed. For the 300 ng/mL cutoff, at 300 ng/mL, 44/76 (qualitative) and 42/77 (semi-quantitative) were observed.
- Spike Recovery: Performed with at least 21 replicates. Qualitative data for 150 ng/mL cutoff showed 25 Negative results at 112.5 ng/mL and 25 Positive results at 187.5 ng/mL. For 300 ng/mL cutoff, 25 Negative results at 225 ng/mL and 25 Positive results at 375 ng/mL were observed.
- Analytical Recovery and Linearity: Demonstrated by spiking drug-free urine to 1000 ng/mL with Benzoylecgonine and diluting to 9 intermediate levels. Each sample run in replicates of five in semi-quantitative mode. Percent recovery ranged from 95.7% to 111.4%.
- Method Comparison and Accuracy: At least 100 patient samples compared against LC-MS/MS.
- Semi-Quantitative Mode Accuracy (150 ng/mL cutoff): Agreement among Positives: 51/51 = 100%. Agreement among Negatives: 49/50 = 98%. One discordant result (Positive by DRI, Negative by LC-MS/MS at 134 ng/mL).
- Semi-Quantitative Mode Accuracy (300 ng/mL cutoff): Agreement among Positives: 50/50 = 100%. Agreement among Negatives: 48/50 = 96%. Two discordant results (Positive by DRI, Negative by LC-MS/MS at 268 ng/mL and 211 ng/mL).
- Qualitative Accuracy (150 ng/mL cutoff): Agreement among Positives: 51/51 = 100%. Agreement among Negatives: 49/50 = 98%. One discordant result (Positive by DRI, Negative by LC-MS/MS at 134 ng/mL).
- Qualitative Accuracy (300 ng/mL cutoff): Agreement among Positives: 50/50 = 100%. Agreement among Negatives: 48/50 = 96%. Two discordant results (Positive by DRI, Negative by LC-MS/MS at 268 ng/mL and 211 ng/mL).
- Specificity: Cross-reactivity of Cocaine and its metabolites and structurally unrelated compounds evaluated. Generally low cross-reactivity for other cocaine metabolites (0.17% to 0.6% for cocaine, cocaethylene, ecgonine, ecgonine methyl ester, norcocaine) except m-hydroxybenzoylecgonine (50%). Structurally unrelated compounds did not show significant cross-reactivity.
- Interference: Effects of pH, endogenous, and exogenous physiologic substances assessed. Controls were accurately detected in presence of interfering substances: Acetaminophen, Acetone, Ascorbic Acid, Aspirin, Caffeine, Creatinine, Ethanol, Galactose, y-Globulin, Glucose, Hemoglobin, Human Serum Albumin, Ibuprofen, Oxalic Acid, Riboflavin, Sodium Chloride, Urea, and pH values from 3 to 11.
- Specific Gravity: Drug-free urine samples within 1.004 to 1.029 spiked with Benzoylecgonine at low and high control concentrations. Controls were accurately detected, indicating no interference.
Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)
- Semi-Quantitative Mode Accuracy (150 ng/mL cutoff): Agreement among Positives: 100%, Agreement among Negative: 98%.
- Semi-Quantitative Mode Accuracy (300 ng/mL cutoff): Agreement among Positives: 100%, Agreement among Negative: 96%.
- Qualitative Accuracy (150 ng/mL cutoff): Agreement among Positives: 100%, Agreement among Negative: 98%.
- Qualitative Accuracy (300 ng/mL cutoff): Agreement among Positives: 100%, Agreement among Negative: 96%.
Predicate Device(s)
Reference Device(s)
Not Found
Predetermined Change Control Plan (PCCP) - All Relevant Information
Not Found
§ 862.3250 Cocaine and cocaine metabolite test system.
(a)
Identification. A cocaine and cocaine metabolite test system is a device intended to measure cocaine and a cocaine metabolite (benzoylecgonine) in serum, plasma, and urine. Measurements obtained by this device are used in the diagnosis and treatment of cocaine use or overdose.(b)
Classification. Class II (special controls). A cocaine and cocaine metabolite test system is not exempt if it is intended for any use other than employment or insurance testing or is intended for Federal drug testing programs. The device is exempt from the premarket notification procedures in subpart E of part 807 of this chapter subject to the limitations in § 862.9, provided the test system is intended for employment and insurance testing and includes a statement in the labeling that the device is intended solely for use in employment and insurance testing, and does not include devices intended for Federal drug testing programs (e.g., programs run by the Substance Abuse and Mental Health Services Administration (SAMHSA), the Department of Transportation (DOT), and the U.S. military).
0
Image /page/0/Picture/0 description: The image contains the logo of the U.S. Food and Drug Administration (FDA). On the left is the Department of Health & Human Services logo. To the right of that is the FDA logo, which is a blue square with the letters "FDA" in white. To the right of the FDA logo is the text "U.S. FOOD & DRUG ADMINISTRATION" in blue.
September 24, 2021
Microgenics Corporation Nikhita Tandon Regulatory Affairs Specialist III 46500 Kato Road Fremont, California 94538
Re: K211973
Trade/Device Name: DRI Cocaine Metabolite Assay Regulation Number: 21 CFR 862.3250 Regulation Name: Cocaine and cocaine metabolite test system Regulatory Class: Class II Product Code: DIO Dated: June 22, 2021 Received: June 25, 2021
Dear Nikhita Tandon:
We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.
If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.
Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal
1
statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801); medical device reporting of medical device-related adverse events) (21 CFR 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safety-reportingcombination-products); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.
Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.
For comprehensive regulatory information about medical devices and radiation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).
Sincerely,
Marianela Perez-Torres, Ph.D. Deputy Director Division of Chemistry and Toxicology Devices OHT7: Office of In Vitro Diagnostics and Radiological Health Office of Product Evaluation and Quality Center for Devices and Radiological Health Food and Drug Administration
Enclosure
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Indications for Use
510(k) Number (if known) K211973
Device Name DRI Cocaine Metabolite Assay
Indications for Use (Describe)
The Alinity c Cocaine assay is a homogeneous enzyme immunoassay intended for the qualitative and/or semi-quantitative determination of benzoylecgonine (Cocaine Metabolite) in human urine at a cutoff concentration of either 150 ng/mL or 300 ng/mL on the Alinity c analyzer.
The semiquantitative application is for the purpose of enabling laboratories to determine an appropriate dilution of the specimen for confirmation by a confirmatory method such as Liquid Chromatography/Tandem Mass Spectrometry (LC-MS/MS) or permitting laboratories to establish quality control procedures.
The assay provides only a preliminary analytical test result. A more specific alternative chemical method must be used to obtain a confirmed analytical result. Gas Chromatography/Mass Spectrometry (GC/MS) or Liquid Chromatography/ Tandem Mass Spectrometry (LC-MS/MS) are the preferred confirmatory method. Tests for cocaine metabolite cannot distinguish between abused drugs and certain prescribed medications.
Clinical and professional judgment should be applied to any drug of abuse test result, particularly when preliminary results are used. For In Vitro Diagnostic Use Only.
| Type of Use (Select one or both, as applicable) | ||||
|---|---|---|---|---|
| > Prescription Use (Part 21 CFR 801 Subpart D) | Over-The-Counter Use (21 CFR 801 Subpart C) |
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3
510(k) Summary
510(k)#: K211973
This 510(k) Summary of Safety and Effectiveness is being submitted in accordance with the requirements of Safe Medical Device Act of 1990 and 21 CFR 807.92.
| A. Device Information | ||||
|---|---|---|---|---|
| Category | Comments |
|---|---|
| Sponsor: | Microgenics Corporation |
| Thermo Fisher Scientific | |
| 46500 Kato Road | |
| Fremont, CA 94538 | |
| Phone: 510-979-5000 | |
| FAX: 510-979-5002 | |
| Correspondent Contact | |
| Information: | Nikhita Tandon |
| Regulatory Affairs Specialist III | |
| Email: nikhita.tandon@thermofisher.com | |
| Phone: 510-979-5000 | |
| Cell:678-964-5599 | |
| FAX: 510-979-5002 | |
| Device Common Name: | Cocaine Metabolite Enzyme Immunoassay |
| Trade or Proprietary Name | DRI Cocaine Metabolite Assay |
| Brand Name | Alinity c Cocaine Assay |
| Predicate Device Product | |
| Code, Classification, | |
| Classification Name & Panel | DIO, Class II, 21 CFR 862. 3250 – Opiate test |
| system, 91 - Toxicology |
Predicate Device Information:
| Predicate Device: | DRI Cocaine Metabolite Assay |
|---|---|
| Predicate Device Manufacturer: | Microgenics Corporation |
| Predicate Device Common Name | Cocaine Metabolite Enzyme Immunoassay |
| Predicate Device Premarket Notification #: | K181499 |
| Predicate Device Product Code, Classification, Classification Name & Panel | DIO, Class II, 21 CFR 862. 3250 – Opiate test system, 91 – Toxicology |
B. Date Summary Prepared
September 16, 2021
C. Description of Device
The DRI Cocaine Metabolite Assay is a homogeneous enzyme immunoassay using ready-touse liquid reagents. The assay uses a specific antibody, which can detect benzoylecgonine in
4
urine. The assay is based on the competition of an enzyme glucose-6-phosphate dehydrogenase (G6PDH) labeled drug and the drug from the urine samplefor a fixed amount of specific antibody binding sites. In the presence of free drug fromthe sample, the free drug occupies the antibody binding sites, allowing the drug-labeled G6PDH to interact with the substrate, resulting in enzyme activity. In the absence of drug from the sample, the specific antibody binds to the drug labeled with G6PDH and the enzyme activity is inhibited. This phenomenon creates a direct relationship between the drug concentration in the urine and the enzyme activity. The enzyme G6PDH activity is determined spectrophotometrically at 340 nm by measuring its ability to convert nicotinamide adenine dinucleotide (NAD) to NADH.
The assay consists of reagents (A and E).
Reagent A: Contains mouse monoclonal anti-benzoylecgonine antibody, glucose-6- phosphate (G6P), and nicotinamide adenine dinucleotide (NAD) in Tris buffer with sodium azide as preservative.
Reagent E: Contains benzoylecgonine analog labeled with glucose-6-phosphate dehydrogenase (G6PDH) in HEPES buffer with sodium azide as preservative.
D. Intended Use
The Alinity c Cocaine assay is a homogeneous enzyme immunoassay intended for the qualitative and/or semi-quantitative determination of benzoylecgonine (Cocaine Metabolite) in human urine at a cutoff concentration of either 150 ng/mL or 300 ng/mL on the Alinity c analyzer.
The semiquantitative application is for the purpose of enabling laboratories to determine an appropriate dilution of the specimen for confirmation by a confirmatory method such as Liquid Chromatography/Tandem Mass Spectrometry (LC-MS/MS) or permitting laboratories to establish quality control procedures.
The assay provides only a preliminary analytical test result. A more specific alternative chemical method must be used to obtain a confirmed analytical result. Gas Chromatography/Mass Spectrometry (GC/MS) or Liquid Chromatography/Tandem Mass Spectrometry (LC-MS/MS) are the preferred confirmatory method. Tests for cocaine metabolite cannot distinguish between abused drugs and certain prescribed medications.
Clinical and professional judgment should be applied to any drug of abuse test result, particularly when preliminary results are used. For In Vitro Diagnostic Use Only.
5
| Characteristics | Candidate Device: DRI
Cocaine Metabolite Assay | Predicate Device: DRI Cocaine
Metabolite Assay(K181499) |
|----------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Intended Use | The Alinity c Cocaine assay is a
homogeneous enzyme immunoassay
intended for the qualitative and/or
semi-quantitative determination of
benzoylecgonine (Cocaine
Metabolite) in human urine at a
cutoff concentration of either 150
ng/mL or 300 ng/mL on the Alinity c
analyzer.
The semiquantitative application is
for the purpose of enabling
laboratories to determine an
appropriate dilution of the specimen
for confirmation by a confirmatory
method such as Liquid
Chromatography/Tandem Mass
Spectrometry (LC-MS/MS) or
permitting laboratories to establish
quality control procedures.
The assay provides only a
preliminary analytical test result. A
more specific alternative chemical
method must be used to obtain a
confirmed analytical result. Gas
Chromatography/Mass Spectrometry
(GC/MS) or Liquid
Chromatography/Tandem Mass
Spectrometry (LC-MS/MS) are the
preferred confirmatory method. Tests
for cocaine metabolite cannot
distinguish between abused drugs
and certain prescribed medications.
Clinical and professional judgment
should be applied to any drug of
abuse test result, particularly when
preliminary results are used. For In
Vitro Diagnostic Use Only. | The DRI Cocaine Metabolite Enzyme
Immunoassay is a homogeneous
enzyme immunoassay intended for
the qualitative and/or semi-
quantitative determination of
benzoylecgonine (Cocaine
Metabolite) in human urine at a
cutoff concentration of either 150
ng/mL or 300 ng/mL.
The semi-quantitative mode is for the
purpose of enabling laboratories to
determine an appropriate dilution of
the specimen for confirmation by a
confirmatory method such as Liquid
Chromatography/tandem mass
spectrometry (LC-MS/MS) or
permitting laboratories to establish
quality control procedures.
The assay provides only a
preliminary analytical test result. A
more specific alternative chemical
method must be used to obtain a
confirmed analytical result. Gas
chromatography / Mass spectrometry
(GC/MS) or Liquid
chromatography/tandem mass
spectrometry (LC-MS/MS) is the
preferred confirmatory method. Tests
for cocaine metabolite cannot
distinguish between abused drugs and
certain prescribed medications.
Clinical and professional judgment
should be applied to any drug of
abuse test result, particularly when
preliminary results are used. For In
Vitro Diagnostic Use Only. |
| Characteristics | Candidate Device: DRI Cocaine
Metabolite Assay | Predicate Device: DRI Cocaine
Metabolite Assay(K181499) |
| Operating
Principle
(Technology) | DRI | Same |
| Measured Analyte | Benzoylecgonine | Same |
| Test Matrix | Urine | Same |
| Cut-off Levels | 150 ng/mL and 300 ng/mL | Same |
| Methodology | Homogeneous Enzyme
Immunoassay | Same |
| Reagents Form | Liquid ready-to-use. | Same |
| Antibody | Mouse monoclonal antibodies | Same |
| Storage | 2–8 °C until expiration date. | Same |
| Principal Operator | Trained professionals | Same |
| Calibrator Levels
for Semi-Quant | 5-point Calibrator | 5-point Calibrator |
| Reference
Instrument | Alinity c Analyzer System | Beckman Coulter AU680 Clinical
Chemistry Analyzer |
E. Comparison to Predicate Device
6
F. Test Principle
The DRI Cocaine Metabolite Assay is a homogeneous enzyme immunoassay using ready-to-use liquid reagents. The assay uses a specific antibody, which can detect benzoylecgonine in urine. The assay is based on the competition of an enzyme glucose-6-phosphate dehydrogenase (G6PDH) labeled drug and the drug from the urine sample for a fixed amount of specific antibody binding sites. In the presence of free drug from the sample, the free drug occupies the antibody binding sites, allowingthe drug-labeled G6PDH to interact with the substrate, resulting in enzyme activity. In the absence of drug from the sample, the specific antibody binds to the drug labeled with G6PDH and the enzyme activity is inhibited. This phenomenon creates a direct relationship between the drug concentration in the urine and the enzyme activity. Theenzyme G6PDH activity is determined spectrophotometrically at 340 nm by measuring its ability to convert nicotinamide adenine dinucleotide (NAD) to NADH.
G. Summary of Supporting Data
1. Analytical Performance:
Performance is evaluated on the Alinity c Analyzer System.
a) Precision
Precision studies were performed in accordance with CLSI Guideline EP05-A3. Samples were prepared by spiking Benzoylecgonine (Cocaine Metabolite) into drug free urine at the cutoff, 25%, 50%, 75% and 100% above and below the cutoff and tested in both qualitative and semi-quantitative
7
modes. Results presented below were generated by testing all samples in replicates of 3, twice per day for20 days, total n=120. The results for both cutoffs are summarized in the tables below:
| Spiked Concentration
(ng/mL) | % of Cutoff
(150 ng/mL) | Total Precision (n=120) | |
|---------------------------------|----------------------------|-------------------------|--------------------------------------------|
| | | # of
Determinants | Immunoassay Results
(Negative/Positive) |
| 0 | -100% | 119 | 119/0 |
| 37.5 | -75% | 120 | 120/0 |
| 75 | -50% | 120 | 120/0 |
| 112.5 | -25% | 120 | 120/0 |
| 150 | 100% | 120 | 76/44 |
| 187.5 | +25% | 120 | 0/120 |
| 225 | +50% | 120 | 0/120 |
| 262.5 | +75% | 120 | 0/120 |
| 300 | +100% | 120 | 0/120 |
Qualitative Study Analysis for 150 ng/mL cutoff
Qualitative Study Analysis for 300 ng/mL cutoff
| Spiked Concentration
(ng/mL) | % of Cutoff
(300 ng/mL) | Total Precision (n=120) | |
|---------------------------------|----------------------------|-------------------------|--------------------------------------------|
| | | # of
Determinants | Immunoassay Results
(Negative/Positive) |
| 0 | -100% | 119 | 119/0 |
| 75 | -75% | 120 | 120/0 |
| 150 | -50% | 120 | 120/0 |
| 225 | -25% | 120 | 120/0 |
| 300 | 100% | 120 | 44/76 |
| 375 | +25% | 120 | 0/120 |
| 450 | +50% | 120 | 0/120 |
| 525 | +75% | 120 | 0/120 |
| 600 | +100% | 120 | 0/120 |
8
| Spiked Concentration
(ng/mL) | % of Cutoff
(150 ng/mL) | Total Precision (n=120) | |
|---------------------------------|----------------------------|--------------------------------------------|-------|
| | # of Determinants | Immunoassay Results
(Negative/Positive) | |
| 0 | -100% | 119 | 119/0 |
| 37.5 | -75% | 120 | 120/0 |
| 75 | -50% | 120 | 120/0 |
| 112.5 | -25% | 120 | 120/0 |
| 150 | 100% | 120 | 73/47 |
| 187.5 | +25% | 120 | 0/120 |
| 225 | +50% | 120 | 0/120 |
| 262.5 | +75% | 120 | 0/120 |
| 300 | +100% | 119 | 0/119 |
Semi-Quantitative Study Analysis for 150 ng/mL cutoff
Semi-Quantitative Study Analysis for 300 ng/mL cutoff
| Spiked Concentration
(ng/mL) | % of Cutoff(300
ng/mL) | # of Determinants | Total Precision (n=120)
Immunoassay Results
(Negative/Positive) |
|---------------------------------|---------------------------|-------------------|-----------------------------------------------------------------------|
| 0 | -100% | 119 | 119/0 |
| 75 | -75% | 120 | 120/0 |
| 150 | -50% | 120 | 120/0 |
| 225 | -25% | 120 | 120/0 |
| 300 | 100% | 119 | 42/77 |
| 375 | +25% | 120 | 0/120 |
| 450 | +50% | 120 | 0/120 |
| 525 | +75% | 120 | 0/120 |
| 600 | +100% | 120 | 0/120 |
b) Spike Recovery
The study was performed for at least 21 replicates. This study was carried out by testing spiked samples containing Benzoylecgonine (Cocaine Metabolite) at the cutoff calibrator and control levels. The spiked samples were prepared by spiking Benzoylecgonine (Cocaine Metabolite) into drug free urine. Samples were tested in both qualitative and semi-quantitative mode. The qualitative results for both cutoffs are summarized in the tables below.
9
| Replicates | 112.5 ng/mL(n=25) | 187.5 ng/mL(n=25) |
|---|---|---|
| 1 | Negative | Positive |
| 2 | Negative | Positive |
| 3 | Negative | Positive |
| 4 | Negative | Positive |
| 5 | Negative | Positive |
| 6 | Negative | Positive |
| 7 | Negative | Positive |
| 8 | Negative | Positive |
| 9 | Negative | Positive |
| 10 | Negative | Positive |
| 11 | Negative | Positive |
| 12 | Negative | Positive |
| 13 | Negative | Positive |
| 14 | Negative | Positive |
| 15 | Negative | Positive |
| 16 | Negative | Positive |
| 17 | Negative | Positive |
| 18 | Negative | Positive |
| 19 | Negative | Positive |
| 20 | Negative | Positive |
| 21 | Negative | Positive |
| 22 | Negative | Positive |
| 23 | Negative | Positive |
| 24 | Negative | Positive |
| 25 | Negative | Positive |
| Overlap | No | No |
| Relative to C/O | All 25 below C/O | All 25 above C/O |
Qualitative Data for 150 ng/mL cutoff
Qualitative Data for 300 ng/mL cutoff
| Replicates | 225 ng/mL(n=25) | 375 ng/mL(n=25) |
|---|---|---|
| 1 | Negative | Positive |
| 2 | Negative | Positive |
| 3 | Negative | Positive |
| 4 | Negative | Positive |
| 5 | Negative | Positive |
| 6 | Negative | Positive |
| 7 | Negative | Positive |
| 8 | Negative | Positive |
| 9 | Negative | Positive |
| 10 | Negative | Positive |
10
| Replicates | 225 ng/mL(n=25) | 375 ng/mL(n=25) |
|---|---|---|
| 11 | Negative | Positive |
| 12 | Negative | Positive |
| 13 | Negative | Positive |
| 14 | Negative | Positive |
| 15 | Negative | Positive |
| 16 | Negative | Positive |
| 17 | Negative | Positive |
| 18 | Negative | Positive |
| 19 | Negative | Positive |
| 20 | Negative | Positive |
| 21 | Negative | Positive |
| 22 | Negative | Positive |
| 23 | Negative | Positive |
| 24 | Negative | Positive |
| 25 | Negative | Positive |
| Overlap | No | No |
| Relative to C/O | All 25 below C/O | All 25 above C/O |
c) Analytical Recovery and Linearity
Linearity studies were performed in accordance with CLSI Guideline EP06-A. To demonstrate the dilution linearity for purposes of sample dilution and quality control of the entire assay range, drug free urine was spiked to the high level calibrator using Benzoylecgonine (Cocaine Metabolite) (1000 ng/mL) and diluted with drug free urine to generate 9 intermediate levels.
Each sample was run in replicates of five in semi-quantitative mode and the average was used to determine percent recovery compared to the expected target value. The percent recovery is summarized in the table below:
| Level | Expected
Concentration(ng/mL) | Observed Mean
Concentration
(ng/mL) | Recovery (%) |
|-------|----------------------------------|-------------------------------------------|--------------|
| 1 | 0.0 | 0.0 | N/A |
| 2 | 103.2 | 98.8 | 95.7 |
| 3 | 206.4 | 230.1 | 111.4 |
| 4 | 309.7 | 344.3 | 111.2 |
| 5 | 412.9 | 428.8 | 103.9 |
| 6 | 516.1 | 502.2 | 97.3 |
| 7 | 619.3 | 655.3 | 105.8 |
| 8 | 722.6 | 757.2 | 104.8 |
| 9 | 825.8 | 892.0 | 108.0 |
| 10 | 929.0 | 922.3 | 99.3 |
| 11 | 1032.2 | 1032.2 | 100.0 |
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d) Method Comparison and Accuracy
The method comparison study was performed in accordance with CLSI Guideline CLSIEP09c 3rd Edition. At least one hundred patient samples were analyzed by the DRI Cocaine Metabolite Assay in both qualitative and semi-quantitative modes and the results were compared to LC-MS/MS. The qualitative and semi-quantitative results for both cutoffs are summarized in the tables below:
Semi-Quantitative Mode Accuracy study with LC-MS/MS as reference method for 150ng/mL cutoff
| Candidate
Device Results | Negativeby
LC-
MS/MS |
225 ng/mL) |
|-----------------------------|----------------------------|-----------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------|---------------------------------------------------------------------------------------|
| Positive | 0 | 0 | 1 | 5 | 46 |
| Negative | 12 | 33 | 4 | 0 | 0 |
Agreement among Positives: 51/51 = 100% Agreement among Negative: 49/50 = 98%
One result was found to be discordant, where the specimen had an interpretation of positive with the DRI Cocaine Metabolite Assay and an interpretation of negative with LC-MS/MS. The discordant results are provided below:
| Sample ID | DRI Cocaine Metabolite Assay | LC-MS/MS | |
|---|---|---|---|
| Interpretation | Benzoylecgonine Concentration | ||
| (ng/mL) | |||
| CEA00125 | Positive | 134 |
Semi-Quantitative Mode Accuracy study with LC-MS/MS as reference method for 300ng/mL cutoff
| Candidate
Device Results | Negative
by LC-
MS/MS |
450 ng/mL) ) |
|-----------------------------|-----------------------------|------------------------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------|
| Positive | 0 | 0 | 2 | 9 | 41 |
| Negative | 12 | 33 | 3 | 0 | 0 |
Agreement among Positives: 50/50 = 100%
Agreement among Negative: 48/50 = 96%
Two results were found to be discordant, where the specimens had an interpretation of positive with the DRI Cocaine Metabolite Assay (Alinity c Cocaine assay) and an interpretation of
12
| DRI Cocaine Metabolite Assay | LC-MS/MS | ||
|---|---|---|---|
| Sample ID | Interpretation | Benzoylecgonine Concentration | |
| (ng/mL) | |||
| CEA00028 | Positive | 268 | |
| CEA00124 | Positive | 211 |
negative with LC-MS/MS. The discordant results are provided below:
Qualitative Accuracy study with LC-MS/MS as reference method for 150 ng/mL cutoff
| Candidate
Device Results | Negative
by LC-
MS/MS |
225 ng/mL) |
|-----------------------------|-----------------------------|-----------------------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------|------------------------------------------------------------------------------------------|
| Positive | 0 | 0 | 1 | 5 | 46 |
| Negative | 12 | 33 | 4 | 0 | 0 |
Agreement among Positives: 51/51 = 100% Agreement among Negative: 49/50 = 98%
One result was found to be discordant, where the specimen had an interpretation of positive with the DRI Cocaine Metabolite Assay and an interpretation of negative with LC-MS/MS. The discordant results are provided below:
| DRI Cocaine Metabolite Assay | LC-MS/MS | |
|---|---|---|
| Sample ID | Interpretation | Benzoylecgonine Concentration |
| (ng/mL) | ||
| CEA00125 | Positive | 134 |
Qualitative Accuracy study with LC-MS/MS as reference method for 300 ng/mL cutoff
| Candidate
Device Results | Negativeby
LC-
MS/MS |
450 ng/mL) |
|-----------------------------|----------------------------|------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------|---------------------------------------------------------------------------------------|
| Positive | 0 | 0 | 2 | 9 | 41 |
| Negative | 12 | 33 | 3 | 0 | 0 |
Agreement among Positives: 50/50 = 100%
Agreement among Negative: 48/50 = 96%
Two results were found to be discordant, where the specimens had an interpretation of positive with the DRI Cocaine Metabolite Assay and an interpretation of negative with LC-MS/MS. The discordant results are provided below:
13
| Sample ID | DRI Cocaine Metabolite Assay
Interpretation | LC-MS/MS
Benzoylecgonine Concentration
(ng/mL) |
|-----------|------------------------------------------------|------------------------------------------------------|
| CEA00028 | Positive | 268 |
| CEA00124 | Positive | 211 |
e) Specificity
The cross-reactivity of Cocaine and its metabolites were evaluated by adding known amounts of each compound to drug-free negative urine. The results are summarized inthe tables below:
| Cocaine and metabolites | Tested Concentration
(ng/mL) | Cross-reactivity(%) |
|--------------------------|---------------------------------|---------------------|
| Benzoylecgonine | 150 | 100 |
| Cocaine | 25,000 | 0.6 |
| Cocaethylene | 30,000 | 0.5 |
| Ecgonine | 90,000 | 0.17 |
| Ecgonine Methyl Ester | 100,000 |