(225 days)
Not Found
No
The device description details a standard enzyme immunoassay based on chemical reactions and spectrophotometric measurement, with no mention of AI or ML algorithms for data analysis or interpretation.
No.
Explanation: The device is an in vitro diagnostic (IVD) assay designed for qualitative and semi-quantitative detection of Hydrocodone and its metabolites in human urine. It is used to provide preliminary analytical test results and is not intended for treatment or mitigation of a disease or condition.
Yes
Explanation: The device is described as an "analytical test" intended for the "qualitative and semi-quantitative detection and estimation of Hydrocodone and its metabolites in human urine." It provides a "preliminary analytical test result" that implies making a finding or determination. This function of detecting and quantifying substances in a biological sample to reveal information about a person's health status (in this case, drug presence) falls under the definition of a diagnostic device.
No
The device is a chemical assay kit comprised of liquid reagents, calibrators, and controls, intended for use with automated clinical chemistry analyzers. It is not a software-only device.
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 semi-quantitative detection and estimation of Hydrocodone and its metabolites in human urine". This is a diagnostic purpose, analyzing a human specimen (urine) to detect the presence of a substance.
- "For In Vitro Diagnostic Use Only": Both the calibrators and controls are labeled with "For In Vitro Diagnostic Use Only". This is a clear indication that the entire system is intended for in vitro diagnostic use.
- Device Description: The description details a laboratory assay that uses reagents to analyze a human sample (urine) outside of the body. This aligns with the definition of an in vitro diagnostic device.
- Performance Studies: The document describes performance studies (Precision, Accuracy, Analytical Recovery and Linearity, Specificity and Cross-Reactivity, Interference, Specific Gravity, Stability) which are typical for evaluating the performance of an IVD.
- Intended User / Care Setting: The assay is designed for "professional use with a number of automated clinical chemistry analyzers," indicating use in a clinical laboratory setting for diagnostic testing.
All of these points strongly support the classification of this device as an In Vitro Diagnostic.
N/A
Intended Use / Indications for Use
DRI® Hydrocodone Assay
The DRI® Hydrocodone Assay is intended for the qualitative and semi-quantitative detection and estimation of Hydrocodone and its metabolites in human urine at a cutoff of 300 ng/mL. The semi-quantitative mode is for purposes of enabling laboratories to determine an appropriate dilution of specimen for confirmatory method such as LC-MS/MS or GC-MS and permitting laboratories to establish quality control measures.
This assay provides a preliminary analytical test result. A more specific alternative chemical method must be used in order to confirm an analytical result. Gas chromatography/mass spectrometry (GC/MS) and Liquid Chromatography/ tandem mass spectrometry (LC-MS/MS) are the preferred confirmatory methods. Clinical consideration and professional judgment should be applied to any drug of abuse test result, particularly when preliminary positive results are used.
DRI® Hydrocodone Calibrators
The DRI® Hydrocodone Assay Calibrators are intended for the DRI® Hydrocodone Assay. For In Vitro Diagnostic Use Only.
DRI® Hydrocodone Controls
The DRI® Hydrocodone Controls are unassayed quality control material intended for use in the DRI Hydrocodone Assay to detect and monitor systematic deviations from accuracy resulting from reagent or instrument defects. For In Vitro Diagnostics Use Only
Product codes (comma separated list FDA assigned to the subject device)
DJG, DLJ, LAS
Device Description
The DRI® Hydrocodone Assay is supplied as a liquid ready-to-use homogeneous enzyme immunoassay. The assay uses specific antibodies that can detect Hydrocodone and its metabolites without any significant cross-reactivity to other opiate compounds. The assay is based on competition between a drug labeled with glucose-6-phosphate dehydrogenase (G6PDH) and free drug from the urine sample, for a fixed amount of specific antibody binding sites. In the absence of free drug from the sample, the specific antibody binds the drug labeled with G6PDH and causes a decrease in enzyme activity. In the presence of free drug, the free drug occupies the antibody binding sites, allowing the drug bound G6PDH to interact with the substrate, resulting in enzyme activity. This phenomenon creates a direct relationship between the drug concentration in urine and enzyme activity. The enzyme activity is determined spectrophotometrically at 340 nm by measuring the conversion of nicotinamide adenine dinucleotide (NAD) to NADH.
The DRI® Hydrocodone Assay is a kit comprised of two reagents, Reagent A and Reagent E, which are bottled separately but sold together within the same kit.
The Reagent A solution contains: mouse monoclonal anti-hydrocodone antibody, glucose-6phosphate (G6P) and nicotinamide adenine dinucleotide (NAD) in Tris buffer with Sodium Azide (≤0.09%) as a preservative). The Reagent E solution contains: glucose-6-phosphate dehydrogenase (G6PDH) in Tris buffer with Sodium Azide (≤0.09%) as preservative.
The DRI® Hydrocodone Enzyme Immunoassay calibrators designated for use at the 300 ng/mL cutoff contain 0 (negative), 100, 300, 500, and 1,000 ng/mL of hydrocodone in human urine matrix with sodium azide (≤0.09%) as preservative. The controls are provided at a concentration of 225 and 375 ng/mL. The calibrators are sold separately and the two controls are sold as a kit.
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
Not Found
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
Not Found
Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)
Precision: 300 ng/mL Cutoff
Samples spiked with various amounts of Hydrocodone were tested in both qualitative and semiquantitative mode using a Clinical Laboratory and Standards Institute (CLSI) protocol. Results presented below were generated by testing all samples in replicates of two, twice per day for 20 days, total n=80.
Qualitative Results using CLSI: All samples tested recovered accurately. Samples at levels below the cutoff read as negative and samples at levels above the cutoff read as positive.
Semi-Quantitative Results Using CLSI: Samples tested recovered accurately. Samples at levels below the cutoff read as negative and samples at levels above the cutoff read as positive.
Accuracy
One hundred patient samples were analyzed by the DRI® Hydrocodone Assay in both qualitative and semi-quantitative modes and the results were compared to the LC-MS/MS. The overall concordance between LC-MS/MS and the DRI Hydrocodone Assay was 93%.
Analytical Recovery and Linearity
To demonstrate linearity for the purpose of sample dilution and quality control over the entire assay range, a drug free urine pool was spiked with Hydrocodone across the range of the calibration curve. Each sample was run in replicates of five on the AU680 instrument in semiquantitative mode and the average was used to determine percent recovery compared to the expected target value. When comparing the result (y) and target (x) value, using the least squares regression technique, the regression equation and correlation are as follows y=1.0341-1.9933 and r2 value was 0.9965. The results demonstrated that the data passed the acceptance criteria. The LC-MS/MS data demonstrated the values were within the acceptance criteria. The results indicated that the dilution linearity exists across the calibrator range.
Specificity and Cross-Reactivity
Several opiates (structurally similar or dissimilar to Hydrocodone) were tested for crossreactivity in the DRI® Hydrocodone Assay. Cross-reactant solutions were prepared by adding known amounts of stock solution to the drug free urine. Hydrocodone, and its metabolites listed in the table were titrated to the lowest levels yielding a positive result in the assay. Structurally related and unrelated opiates listed in the table were tested and results are shown.
Interference
The potential interference of pH and endogenous physiologic substances on recovery of Hydrocodone using DRI® Hydrocodone Assay was assessed by spiking known compounds of potentially interfering substances into the low (225 ng/mL) and high (375 ng/mL) controls for 300 ng/mL cutoff. In the presence of the compounds listed, the controls were detected accurately indicating that these compounds did not show interference in the assay. The samples were tested in both qualitative and semi-quantitative mode.
Specific Gravity
Drug Free urine samples with specific gravity ranging in value from 1.000 to 1.036 g/mL were split and either left unspiked or spiked to a final concentration of either 225 ng/mL or 375 ng/mL (the low and high control concentrations, respectively). These samples were then evaluated in qualitative and semi-quantitative modes. No interference was observed.
Stability:
Open Vial Calibrators and Controls
Open vial stability studies for three lots stored at 2-8°C were carried out to support a claim of 60 days at this time for qualitative and semi-quantitative.
Real Time Stability for Reagent, Calibrators and Controls
Real time studies for reagents, calibrators and controls at 2-8°C are ongoing and have been carried out up to 289 days.
Accelerated Stability Results for Reagents, Calibrators and Controls
Accelerated testing results show that the low control was detected as negative and the high control was detected as positive for each time point for a period of 4 months at 23°C. This is equivalent to 13 months of stability according to Q10 math model. The % recoveries of the low and high control were within 80-120%. The data for six month accelerated stability also confirmed that the low control was detected as negative and the high control was detected as positive.
Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)
Overall concordance between LC-MS/MS and the DRI Hydrocodone Assay was 93%.
Predicate Device(s): If the device was cleared using the 510(k) pathway, identify the Predicate Device(s) K/DEN number used to claim substantial equivalence and list them here in a comma separated list exactly as they appear in the text. List the primary predicate first in the list.
Reference Device(s): Identify the Reference Device(s) K/DEN number and list them here in a comma separated list exactly as they appear in the text.
Not Found
Predetermined Change Control Plan (PCCP) - All Relevant Information for the subject device only (e.g. presence / absence, what scope was granted / cleared under the PCCP, any restrictions, etc).
Not Found
§ 862.3650 Opiate test system.
(a)
Identification. An opiate test system is a device intended to measure any of the addictive narcotic pain-relieving opiate drugs in blood, serum, urine, gastric contents, and saliva. An opiate is any natural or synthetic drug that has morphine-like pharmocological actions. The opiates include drugs such as morphine, morphine glucoronide, heroin, codeine, nalorphine, and meperedine. Measurements obtained by this device are used in the diagnosis and treatment of opiate use or overdose and in monitoring the levels of opiate administration to ensure appropriate therapy.(b)
Classification. Class II (special controls). An opiate 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/1 description: The image shows the logo for the Department of Health & Human Services - USA. The logo consists of a circular seal with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" arranged around the perimeter. Inside the circle is an abstract symbol resembling a stylized eagle or bird with three human profiles incorporated into its design.
Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002
October 9, 2015
MICROGENICS CORPORATION LAURIE WONG MANAGER, REGULATORY AFFAIRS 46500 KATO ROAD FREMONT CA 94538
Re: K150502
Trade/Device Name: DRI Hydrocodone Assay, DRI Hydrocodone Calibrators, DRI Hydrocodone Controls Regulation Number: 21 CFR 862.3650 Regulation Name: Opiate Test System Regulatory Class: II Product Code: DJG, DLJ, LAS Dated: August 14, 2015 Received: August 19, 2015
Dear Ms. Wong:
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. 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 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 Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the
1
electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.
If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 809), please contact the Division of Industry and Consumer Education at its toll-free number (800) 638 2041 or (301) 796-7100 or at its Internet address
http://www.fda.gov/MedicalDevices/Resourcesfor You/Industry/default.htm. 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
http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.
You may obtain other general information on your responsibilities under the Act from the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address
http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.
Sincerely yours,
Courtney
Courtney H. Lias, Ph.D. Director Division of Chemistry and Toxicology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health
Enclosure
2
Indications for Use
510(k) Number (if known) K150502
Device Name DRI® Hydrocodone Assay DRI® Hydrocodone Calibrators DRI® Hydrocodone Controls
Indications for Use (Describe)
DRI® Hydrocodone Assay
The DRI® Hydrocodone Assay is intended for the qualitative and semi-quantitative detection and estimation of Hydrocodone and its metabolites in human urine at a cutoff of 300 ng/mL. The semi-quantitative mode is for purposes of enabling laboratories to determine an appropriate dilution of specimen for confirmatory method such as LC-MS/MS or GC-MS and permitting laboratories to establish quality control measures.
This assay provides a preliminary analytical test result. A more specific alternative chemical method must be used in order to confirm an analytical result. Gas chromatography/mass spectrometry (GC/MS) and Liquid Chromatography/ tandem mass spectrometry (LC-MS/MS) are the preferred confirmatory methods. Clinical consideration and professional judgment should be applied to any drug of abuse test result, particularly when preliminary positive results are used.
DRI® Hydrocodone Calibrators
The DRI® Hydrocodone Assay Calibrators are intended for the DRI® Hydrocodone Assay. For In Vitro Diagnostic Use Only.
DRI® Hydrocodone Controls
The DRI® Hydrocodone Controls are unassayed quality control material intended for use in the DRI Hydrocodone Assay to detect and monitor systematic deviations from accuracy resulting from reagent or instrument defects. For In Vitro Diagnostics 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 of Safety and Effectiveness
This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of SMDA 1990 and 21 CFR 807.92.
Introduction
According to the requirements of 21 CFR 807.92, the following information provides sufficient details to understand the basis for a determination of substantial equivalence.
Submitter Name, Address and Content:
Microgenics 46500 Kato Road Fremont, CA 94538 Phone: (510) 979-5000 Fax: (510) 979-5002 Email: laurie.wong@thermofisher.com
Contact: Laurie Wong Regulatory Affairs Manager
Date Summary Originally Prepared: February 25, 2015 Date Summary updated per FDA request of October 1, 2015
Device Name and Classification
Trade or Proprietary Name:
DRI® Hydrocodone Assay DRI® Hydrocodone Assay Calibrators DRI® Hydrocodone Assay Controls
Common Name: Homogeneous Hydrocodone Enzyme Immunoassay
| Classification Name: | Enzyme Immunoassay, Hydrocodone |
|---|---|
| Class II, DJG (91 Toxicology) | |
| 21 CFR 862.3650 |
Drug Specific Calibrators Class II, DLJ (91 Toxicology) 21 CFR 862.3200
Drug Specific Controls Class I, Reserved LAS (91 Toxicology) 21 CFR 862.3280
4
Legally Marketed Predicate Device(s)
The DRI® Hydrocodone Assay is substantially equivalent to the predicate Lin-Zhi International, Inc., LZI Hydrocodone Enzyme Immunoassay (K141055). The DRI® Hydrocodone Assay, calibrators, and controls are identical or similar to the predicate in terms of intended use, method principle, device components, and clinical performance.
Device Description
The DRI® Hydrocodone Assay is supplied as a liquid ready-to-use homogeneous enzyme immunoassay. The assay uses specific antibodies that can detect Hydrocodone and its metabolites without any significant cross-reactivity to other opiate compounds. The assay is based on competition between a drug labeled with glucose-6-phosphate dehydrogenase (G6PDH) and free drug from the urine sample, for a fixed amount of specific antibody binding sites. In the absence of free drug from the sample, the specific antibody binds the drug labeled with G6PDH and causes a decrease in enzyme activity. In the presence of free drug, the free drug occupies the antibody binding sites, allowing the drug bound G6PDH to interact with the substrate, resulting in enzyme activity. This phenomenon creates a direct relationship between the drug concentration in urine and enzyme activity. The enzyme activity is determined spectrophotometrically at 340 nm by measuring the conversion of nicotinamide adenine dinucleotide (NAD) to NADH.
The DRI® Hydrocodone Assay is a kit comprised of two reagents, Reagent A and Reagent E, which are bottled separately but sold together within the same kit.
The Reagent A solution contains: mouse monoclonal anti-hydrocodone antibody, glucose-6phosphate (G6P) and nicotinamide adenine dinucleotide (NAD) in Tris buffer with Sodium Azide (≤0.09%) as a preservative). The Reagent E solution contains: glucose-6-phosphate dehydrogenase (G6PDH) in Tris buffer with Sodium Azide (≤0.09%) as preservative.
The DRI® Hydrocodone Enzyme Immunoassay calibrators designated for use at the 300 ng/mL cutoff contain 0 (negative), 100, 300, 500, and 1,000 ng/mL of hydrocodone in human urine matrix with sodium azide (≤0.09%) as preservative. The controls are provided at a concentration
5
of 225 and 375 ng/mL. The calibrators are sold separately and the two controls are sold as a kit. Intended Use
The DRI® Hydrocodone Assay is intended for the qualitative and semi-quantitative detection and estimation of Hydrocodone and its metabolites in human urine at a cutoff of 300 ng/mL. The semi-quantitative mode is for purposes of enabling laboratories to determine an appropriate dilution of specimen for confirmation by a confirmatory method such as LC-MS/MS or GC-MS and permitting laboratories to establish quality control measures.
This assay provides a preliminary analytical test result. A more specific alternative chemical method must be used in order to confirm an analytical result. Gas chromatography/mass spectrometry (GC/MS) and Liquid Chromatography/tandem mass spectrometry (LC-MS/MS) are the preferred confirmatory methods. Clinical consideration and professional judgment should be applied to any drug of abuse test result, particularly when preliminary positive results are used.
The DRI® Hydrocodone Assay Calibrators are intended for the calibration of the DRI® Hydrocodone Assay. For In Vitro Diagnostic Use Only.
The DRI® Hydrocodone Controls are unassayed quality control material intended for use in the DRI Hydrocodone Assay to detect and monitor systematic deviations from accuracy resulting from reagent or instrument defects. For In Vitro Diagnostics Use Only
Comparison to Predicate Device
The DRI® Hydrocodone Assay is substantially equivalent to the predicate Lin-Zhi International, Inc., LZI Hydrocodone Enzyme Immunoassay, Calibrators and Controls cleared by the FDA under the premarket notification K141055 for its stated intended use.
The following table (next page) compares the DRI® Hydrocodone Assay to the predicate device.
6
| Device | Subject Device | Predicate Device |
|---|---|---|
| Characteristics | DRI Hydrocodone Assay, Calibrators, and Controls | LZI Hydrocodone Enzyme Immunoassay, Calibrators and Controls (K141055) |
| Intended Use | The DRI® Hydrocodone Assay is | |
| intended for the qualitative and | ||
| semi-quantitative detection | ||
| and estimation of Hydrocodone and | ||
| its metabolites in human urine at a | ||
| cutoff of 300 ng/mL. The | ||
| semi-quantitative mode is for | ||
| purposes of enabling laboratories to | ||
| determine an appropriate | ||
| dilution of specimen for | ||
| confirmation by a confirmatory | ||
| method such as LC-MS/MS or GC- | ||
| MS and permitting laboratories to | ||
| establish quality control measures. |
This assay provides a preliminary
analytical test result. A more
specific alternative chemical
method must be used in order to
confirm an analytical result. Gas
chromatography/mass
spectrometry (GC/MS) and Liquid
Chromatography/tandem mass
spectrometry (LC-MS/MS)
are the preferred confirmatory
methods. Clinical consideration and
professional judgment
should be applied to any drug of
abuse test result, particularly when
preliminary positive results are used.
The DRI® Hydrocodone Assay
Calibrators are intended for the
calibration of the DRI®
Hydrocodone Assay. The DRI®
Hydrocodone Controls are used to
validate the DRI® Hydrocodone
Assay calibration. For In Vitro
Diagnostics Use Only. | The LZI Hydrocodone Enzyme
Immunoassay, when used in
conjunction with the AU480
automated clinical system analyzers,
is intended for the qualitative and
semi-quantitative determination of
hydrocodone in human urine at
cutoff values of 100 or 300 ng/mL.
The assay is designed for
professional use with a number of
automated clinical chemistry
analyzers.
This assay provides a rapid
screening procedure for determining
the presence of hydrocodone and
hydromorphone in urine. The assay
provides only a preliminary
analytical result. A more specific
alternative chemical method must be
used in order to obtain a confirmed
analytical result Gas or liquid
chromatography/mass spectrometry
(GC/MS or LC/MS) is the preferred
confirmatory method. Clinical
consideration and professional
judgment should be exercised with
any drug of abuse test result,
particularly when the preliminary
test result is positive. |
| Analyte | Hydrocodone | Hydrocodone |
| Cutoff Level | 300 ng/mL | 100 or 300 ng/mL |
| Sample Matrix | Human Urine Matrix | Human Urine Matrix |
| Calibrators Level | 300 ng/mL Cutoff: 5 levels: 0, 100,
300, 500 and 1,000 ng/mL | 100 ng/mL Cutoff: 5 levels: 0, 50,
100, 150, and 300 ng/mL
300 ng/mL Cutoff: 5 levels: 0, 150,
300, 500, and 800 ng/mL |
| Controls Level | 300 ng/mL Cutoff: 2 Levels (225
ng/mL and 375 ng/mL) | 100 ng/mL Cutoff: 2 Levels (75
ng/mL, 125 ng/mL) |
| Test Principle | The DRI® Hydrocodone Assay is
supplied as a liquid ready-to-use
homogeneous enzyme
immunoassay. The assay uses
specific antibodies that can detect
Hydrocodone without any
significant cross-reactivity to other
opiate compounds. The assay is
based on competition between a
drug labeled with glucose-6-
phosphate dehydrogenase (G6PDH)
and free drug from the urine sample,
for a fixed amount of specific
antibody binding sites. In the
absence of free drug from the
sample, the specific antibody binds
the drug labeled with G6PDH and
causes a decrease in enzyme
activity. This phenomenon creates a
direct relationship between the drug
concentration in urine and enzyme
activity. The enzyme activity is
determined spectrophotometrically
at 340 nm by measuring the
conversion of nicotinamide adenine
dinucleotide (NAD) to NADH. | The LZI Hydrocodone Enzyme
Immunoassay is a liquid ready-to-
use homogeneous enzyme
immunoassay. The assay uses
specific antibodies that can detect
Hydrocodone without any
significant cross-reactivity to other
opiate compounds. |
| Storage | 2-8°C until expiration date | 2-8°C until expiration date |
7
8
Performance Characteristics Summary:
AU680 Analyzer
Precision: 300 ng/mL Cutoff
Samples spiked with various amounts of Hydrocodone were tested in both qualitative and semiquantitative mode using a Clinical Laboratory and Standards Institute (CLSI) protocol. Results presented below were generated by testing all samples in replicates of two, twice per day for 20 days, total n=80.
| Within Run Precision (n=80) | Total Run Precision (n=80) | |||||
|---|---|---|---|---|---|---|
| Hydrocodone | ||||||
| Spike | ||||||
| Concentration | ||||||
| (ng/mL) | % of | |||||
| Cutoff | LC- | |||||
| MS/MS | ||||||
| (ng/mL) | Number of | |||||
| determinants | Immunoassay | |||||
| Results | Number of | |||||
| determinants | Immunoassay | |||||
| Results | ||||||
| 0 | -100% | 0 | 80 | 80 Neg | 80 | 80 Neg |
| 75 | -75% | 87 | 80 | 80 Neg | 80 | 80 Neg |
| 150 | -50% | 171 | 80 | 80 Neg | 80 | 80 Neg |
| 225 | -25% | 255 | 80 | 80 Neg | 80 | 80 Neg |
| 300 | 100% | 345 | 80 | 46 Neg/34 Pos | 80 | 46 Neg/34 Pos |
| 375 | +25% | 442 | 80 | 80 Pos | 80 | 80 Pos |
| 450 | +50% | 535 | 80 | 80 Pos | 80 | 80 Pos |
| 525 | +75% | 561 | 80 | 80 Pos | 80 | 80 Pos |
| 600 | +100% | 664 | 80 | 80 Pos | 80 | 80 Pos |
Qualitative Results using CLSI:
All samples tested recovered accurately. Samples at levels below the cutoff read as negative and samples at levels above the cutoff read as positive.
Semi-Quantitative Results Using CLSI:
| Within Run Precision (n=80) | Total Run Precision (n=80) | |||||
|---|---|---|---|---|---|---|
| Hydrocodone | ||||||
| Spike | ||||||
| Concentration | ||||||
| (ng/mL) | % of | |||||
| Cutoff | LC- | |||||
| MS/MS | ||||||
| (ng/mL) | Number of | |||||
| determinants | Immunoassay | |||||
| Results | Number of | |||||
| determinants | Immunoassay | |||||
| Results |
9
| 0 | -100% | 0 | 80 | 80 Neg | 80 | 80 Neg |
|---|---|---|---|---|---|---|
| 75 | -75% | 87 | 80 | 80 Neg | 80 | 80 Neg |
| 150 | -50% | 171 | 80 | 80 Neg | 80 | 80 Neg |
| 225 | -25% | 255 | 80 | 80 Neg | 80 | 80 Neg |
| 300 | 100% | 345 | 80 | 40 Neg/40 Pos | 80 | 40 Neg/40 Pos |
| 375 | +25% | 442 | 80 | 80 Pos | 80 | 80 Pos |
| 450 | +50% | 535 | 80 | 80 Pos | 80 | 80 Pos |
| 525 | +75% | 561 | 80 | 80 Pos | 80 | 80 Pos |
| 600 | +100% | 664 | 80 | 80 Pos | 80 | 80 Pos |
Accuracy
One hundred patient samples were analyzed by the DRI® Hydrocodone Assay in both qualitative and semi-quantitative modes and the results were compared to the LC-MS/MS. The overall concordance between LC-MS/MS and the DRI Hydrocodone Assay was 93%.
Qualitative Results with LC-MS/MS as reference method
| DRI
Hydroco-
done
Assay | Negative | Less than
50% of
cutoff
concentrat
ion by
LC/MS
analysis
(450
ng/mL |
|----------------------------------|----------|-----------------------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------|------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------------------|
| Positive | 0 | 1* | 5** | 10 | 39 |
| Negative | 31 | 6 | 7 | 1 | 0 |
10
*and ** 1 Sample in each bin has oxycodone concentrations greater than >37,000 ng/mL
| DRI
Hydroco-
done
Assay | Negative | Less than
50% of
cutoff
concentrat
ion by
Predicate
analysis
(450
ng/mL |
|----------------------------------|----------|---------------------------------------------------------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------------------|
| Positive | 0 | 1* | 5** | 10 | 39 |
| Negative | 31 | 6 | 7 | 1 | 0 |
Semi-Quantitative Results with LC-MS/MS as reference method
*and ** 1 Sample in each bin has oxycodone concentrations greater than >37,000 ng/mL
Discordant Result Table for the Discrepant Samples near cutoff
| Sample # | Qualitative EIA | LC-MS/MS (ng/mL) | Semi-Quantitative EIA (ng/mL) | |||
|---|---|---|---|---|---|---|
| Hydrocodone | Hydromorphone | Hydormorphone 3β-D glucuronide | Adjusted Total# | |||
| 33 | Positive | 143.3 | 37,000 ng/mL. |
*LLOO = Lowest Limit of Quantitation is 40 ng/mL
Analytical Recovery and Linearity
To demonstrate linearity for the purpose of sample dilution and quality control over the entire assay range, a drug free urine pool was spiked with Hydrocodone across the range of the calibration curve. Each sample was run in replicates of five on the AU680 instrument in semiquantitative mode and the average was used to determine percent recovery compared to the expected target value. When comparing the result (y) and target (x) value, using the least squares regression technique, the regression equation and correlation are as follows y=1.0341-1.9933 and r2 value was 0.9965.
Linearity Results for Lot 3
| Target
Hydrocodone
(ng/mL) | Observed
Recovery
(ng/mL)
N=5 | Recovery % |
|----------------------------------|----------------------------------------|------------|
| 0 | N/A | N/A |
| 50 | 47 | 94 |
| 75 | 76 | 101 |
| 100 | 108 | 108 |
| 150 | 171 | 114 |
| 225 | 250 | 111 |
| 300 | 302 | 101 |
| 375 | 398 | 106 |
| 450 | 472 | 105 |
| 500 | 527 | 105 |
| 750 | 844 | 113 |
| 1000 | 1014 | 101 |
The results demonstrated that the data passed the acceptance criteria. The LC-MS/MS data demonstrated the values were within the acceptance criteria. The results indicated that the
12
dilution linearity exists across the calibrator range.
Specificity and Cross-Reactivity
Several opiates (structurally similar or dissimilar to Hydrocodone) were tested for crossreactivity in the DRI® Hydrocodone Assay. Cross-reactant solutions were prepared by adding known amounts of stock solution to the drug free urine.
Hydrocodone, and its metabolites listed in the table were titrated to the lowest levels yielding a positive result in the assay.
Structurally related and unrelated opiates listed in the table were tested and results are shown below.
| Hydrocodone and its
metabolites | Tested
Concentration
(ng/mL) | Pos/Neg | % Cross-
reactivity |
|------------------------------------|------------------------------------|---------|------------------------|
| Neg urine | 0 | Pos | 0% |
| Hydrocodone | 300 | Pos | 102% |
| Hydromorphone | 250 | Pos | 122% |
| Hydromorphone-3β-
glucuronide | 250 | Pos | 122% |
| Norhydrocodone | 10,000 | Pos | 3.1% |
| Dihydrocodeine | 11,000 | Pos | 2.7% |
Cross-Reactivity Results for Lot 3
| Structurally related
compounds and other
opiates | Tested
Concentration
(ng/mL) | Pos/Neg | % Cross-
reactivity |
|--------------------------------------------------------|------------------------------------|---------|------------------------|
| 6-Acetyl Morphine | 100,000 | Neg |