(123 days)
An Enzyme Immunoassay for the in vitro diagnostic quantitative measurement of free active testosterone in saliva. Measurement of testosterone is used in the diagnosis and treatment of disorders involving the male sex hormones (androgens), including primary and secondary hypogonadism, delaved or precocious puberty, impotence in males and, in females hirsutism (excessive hair) and virilization (masculinization) due to tumors, polycystic ovaries, and adrenogenital syndromes.
The DRG Salivary Testosterone ELISA Kit is based on the competition princible and the microplate separation. An unknown amount of free testosterone present in the sample and a fixed amount of testosterone conjugated with horseradish peroxidase compete for the binding sites of mouse monoclonal testosterone -antiserum coated onto the wells. After one hour incubation the microplate is washed to stop the competition reaction. After addition of the substrate solution the concentration of testosterone is inversely proportional to the optical density measured.
The DRG Salivary Testosterone ELISA is a diagnostic device used for the quantitative measurement of free-active testosterone in saliva. The device performance was evaluated through various studies, including method comparison, sensitivity, specificity, reproducibility, recovery, and linearity.
1. Table of Acceptance Criteria and Reported Device Performance
| Performance Metric | Acceptance Criteria (Implicit) | Reported Device Performance |
|---|---|---|
| Normal Range | Establish a 5-95% reference range for apparently healthy adult males and females across different age groups. | Determined from 187 adult males (21-75 years) and 188 adult females (21-75 years). Males (pg/mL):- 21-30: 47.2 - 136.2- 31-40: 46.8 - 106.8- 41-50: 36.5 - 82.7- 51-60: 19.1 - 89.0- 61-75: 12.2 - 68.6 Females (pg/mL):- 21-30: 7.9 - 50.4- 31-40: <7.0 - 44.8- 41-50: <7.0 - 39.4- 51-60: <7.0 - 29.8- 61-75: <7.0 - 29.3 |
| Method Comparison | Demonstrate substantial equivalence to a commercially available LIA method for measuring testosterone in saliva, typically indicated by a strong correlation coefficient and acceptable regression analysis. Consistency in reporting is implicitly expected though specific thresholds are not provided. | Study 1: 99 male and female subjects (20-70 years). - Correlation: 0.904 - Regression: y = 0.9251x - 7.4369 (vs LIA)Study 2: 81 additional saliva samples (40-65 year old men and women). - R² = 0.9866- Regression: y = 1.0057x - 2.4196 (DRG ELISA vs LIA) |
| Sensitivity | The lowest detectable level of testosterone distinguishable from a zero standard (analytical sensitivity) and the lowest functional sensitivity at a specified confidence limit. | - Lowest analytical detectable level: 1.857 pg/mL at 95% confidence limit.- Lowest functional sensitivity: 7.1 pg/mL at 95% confidence limit. |
| Specificity | Minimal cross-reactivity with other related steroids and compounds, particularly those structurally similar, to ensure accurate testosterone measurement. | - Testosterone: 100%- 5α-Dihydrotestosterone: 0.80%- Androstenedione: 0.90%- 11β-hydroxysterone: 3.30%- 17α-methyltestosterone: 0.10%- 19-Nortestosterone: 3.30%- Epitestosterone: 0.10%- Estradiol: 0.10%- Progesterone: < 0.10%- Cortisol: < 0.10% - Estrone: < 0.10%- Danazol: < 0.10% |
| Reproducibility | Demonstrate acceptable variability (CV%) for intra-assay (within-run), inter-assay (between-run), and inter-lot (between-kit-lots) measurements across a range of testosterone concentrations. Typically, CV% values should be low, indicating precision. | Intra-Assay (n=20 replicates/sample per run):- CV% range: 6.23% - 13.81% (across 5 samples with mean range 12.94-144.00 pg/mL)Inter-Assay (n=20 duplicate measurements over 10 days/sample):- CV% range: 5.51% - 9.62% (across 5 samples with mean range 33.61-823.08 pg/mL)Inter-Lot (n=9 triplicate measurements over 3 kit lots/sample):- CV% range: 2.90% - 5.85% (across 5 samples with mean range 44.00-517.65 pg/mL) |
| Recovery | Recover close to 100% of added analyte in spiked samples, demonstrating accuracy and lack of matrix interference. A typical range for acceptable recovery is 90-110%. | Average Recovery: - Range from 92.35% to 104.92% (from 6 saliva samples with various spiked concentrations).Individual Recoveries (selected examples from samples 1-6):- 95.56%, 101.73%, 104.92%, 95.82%- 97.19%, 103.32%, 104.30%, 92.35%- 99.25%, 95.03%, 94.75%, 96.69%- 97.8%, 98.6%, 100.0%- 98.7%, 96.0%, 97.7%- 99.1%, 102.3%, 100.5% |
| Linearity | Demonstrate that the assay provides proportional results across its claimed analytical measurement range, typically assessed by percentage recovery of serially diluted samples. An acceptable range for recovery is generally 90-110%. The functional range should be clearly defined. | Usable Range: 7.1 - 4500 pg/mL.Average % Recovery: 97.5% - 100.8% (for 6 samples with concentrations from 440.00 to 8000.0 pg/mL).Range of Recovery %: - From 93.6% to 107.8% (across 6 samples). (Three native samples were serially diluted, and 3 samples were spiked and then serially diluted up to 1:128). |
2. Sample Sizes Used for the Test Set and Data Provenance:
- Normal Range Study: 187 adult male and 188 adult female apparently healthy subjects (ages 21-75 years). Samples were collected in the morning. Data provenance is not explicitly stated beyond "apparently healthy subjects," but given the domestic contact information (New Jersey) and FDA submission, it implicitly refers to data collected within the US or compliant with US regulatory standards. This appears to be a prospective collection for this study.
- Method Comparison (Study 1): 99 male and female subjects (ages 20-70 years).
- Method Comparison (Study 2): 81 additional saliva samples from 40-65 year old men and women.
- Sensitivity: Not specified as a separate test set, derived from standard curve analysis.
- Specificity: Not specified as a separate test set, derived from testing specific steroids and compounds.
- Reproducibility (Intra-assay): 5 saliva samples, 20 replicate measurements per sample.
- Reproducibility (Inter-assay): 5 saliva samples, duplicate measurements over 10 days per sample.
- Reproducibility (Inter-lot): 5 saliva samples, triplicate measurements in three different kit lots per sample.
- Recovery: 6 different saliva samples.
- Linearity: 6 saliva samples (3 native, 3 spiked).
The data provenance for all studies is not explicitly stated but is implicitly within the context of a US-based manufacturer seeking FDA clearance, suggesting data generated for this purpose. The studies described are likely prospective or specifically conducted for the validation of this device.
3. Number of Experts Used to Establish Ground Truth for the Test Set and Their Qualifications:
This device is an immunoassay for quantitative measurement. The "ground truth" for such devices often refers to the actual concentration of the analyte, established either by:
* Traceability to recognized reference materials/methods.
* Performance against a well-established, validated comparative method (often called a "predicate" device in regulatory submissions, or a "gold standard" method).
In this context, the "ground truth" for the method comparison studies was established by a "commercially available LIA method" (presumably the predicate device or a reference method). No human experts or their qualifications are mentioned for establishing the ground truth for this type of quantitative assay, as the measurement is chemical/analytical.
4. Adjudication Method for the Test Set:
Not applicable. Adjudication methods (like 2+1, 3+1) are typically used for qualitative or semi-quantitative assessments, especially in imaging or clinical diagnosis where human interpretation is involved. For a quantitative immunoassay like this, the result is a numerical value, and "adjudication" in the traditional sense is not performed. Accuracy is determined by comparing measured values to known concentrations or reference method results.
5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study Was Done:
No, an MRMC comparative effectiveness study was not done. This type of study is typically relevant for diagnostic imaging devices or other tools where human interpretation of results is a critical component and the effect of AI on human reader performance is being evaluated. This device is an automated, quantitative immunoassay.
6. If a Standalone Study (i.e., algorithm only without human-in-the-loop performance) Was Done:
Yes, a standalone study was done. All performance metrics described (Method Comparison, Sensitivity, Specificity, Reproducibility, Recovery, Linearity) represent the performance of the DRG SLV Testosterone ELISA device itself, in an automated or semi-automated laboratory setting, without direct human-in-the-loop interpretation impacting the measurement results.
7. The Type of Ground Truth Used:
The primary ground truth used for performance evaluation was:
- Comparative Method: For method comparison studies, the results from a "commercially available LIA method" were used as the reference to assess the DRG device's agreement and substantial equivalence.
- Spiked Samples/Known Concentrations: For sensitivity, recovery, and linearity studies, known concentrations of testosterone (either added to samples or derived from serial dilutions) served as the ground truth.
- Reference Materials: Implicitly, the sensitivity and specificity characterization would rely on well-characterized reference materials or pure chemical compounds.
8. The Sample Size for the Training Set:
This document describes a diagnostic assay kit (ELISA), not an AI algorithm. Therefore, there is no "training set" in the context of machine learning. The studies described are validation studies to characterize the assay's analytical performance.
9. How the Ground Truth for the Training Set Was Established:
Not applicable, as there is no "training set" for an ELISA kit. The "ground truth" (i.e., reference values) for the validation studies were established as described in section 7.
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JAN 2 7 2006
SUMMARY OF SAFETY AND EFFECTIVENESS FOR DRG SALIVARY TESTOSTERONE ELISA
| Manufacturer: | DRG International, Inc.1167 U.S. Highway 22Mountainside, NJ 07092 |
|---|---|
| Contact Information: | Lehnus & AssociatesGary Lehnus150 Cherry Lane Rd.East Stroudsburg, PA 18301Tel: (570) 620-0198 |
Device Name / Classification:
The device trade name is the DRG SLV Testosterone ELISA having FDA assigned name: Testosterone test system, 21 CFR, 862.1680, categorized as Class I "reserved" medical devices for the Clinical Chemistry and Clinical Toxicology Panel, as Product Code CDZ.
Test Principle
The DRG Salivary Testosterone ELISA Kit is based on the competition princible and the microplate separation. An unknown amount of free testosterone present in the sample and a fixed amount of testosterone conjugated with horseradish peroxidase compete for the binding sites of mouse monoclonal testosterone -antiserum coated onto the wells. After one hour incubation the microplate is washed to stop the competition reaction. After addition of the substrate solution the concentration of testosterone is inversely proportional to the optical density measured.
Device Intended Use:
An Enzyme Immunoassay for the in vitro diagnostic quantitative measurement of free active testosterone in saliva. Measurement of testosterone is used in the diagnosis and treatment of disorders involving the male sex hormones (androgens), including primary and secondary hypogonadism, delaved or precocious puberty, impotence in males and, in females hirsutism (excessive hair) and virilization (masculinization) due to tumors, polycystic ovaries, and adrenogenital syndromes.
Device Performance Normal Range Study
In order to determine the normal range of SLV Testosterone, saliva samples from 187 adult male and 188 adult female apparently healthy subjects, ages 21 to 75 years, were collected in the morning and analyzed using the DRG SLV Testosterone ELISA kit. The following range was calculated from this study.
| Men $\male$ | Women $\female$ | |||||
|---|---|---|---|---|---|---|
| Age GroupYears | Range(5 - 95%) | Median | n | Range(5 - 95%) | Median | n |
| 21 - 30 | 47.2 - 136.2 | 92.8 | 42 | 7.9 - 50.4 | 20.8 | 40 |
| 31 - 40 | 46.8 - 106.8 | 73.6 | 37 | <7.0 - 44.8 | 17.1 | 40 |
| 41 - 50 | 36.5 - 82.7 | 58.8 | 34 | <7.0 - 39.4 | 18.3 | 38 |
| 51 - 60 | 19.1 - 89.0 | 44.5 | 36 | <7.0 - 29.8 | 19.2 | 38 |
| 61 - 75 | 12.2 - 68.6 | 38.9 | 38 | <7.0 - 29.3 | 16.0 | 32 |
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Method comparison
A study was performed that evaluated saliva samples from 99 male and female subjects ages 20 to 70 years. The saliva samples were run in duplicate on the DRG test and a commercially available LIA method to determine the concentration of free Testosterone in the samples. A correlation of 0.904 and regression formula of y = 0.9251x - 7.4369 was obtained versus this method.
Another study was performed to further evaluate the substantial equivalence of the DRG SLV Testosterone to the LIA saliva test. The concentration of testosterone in 81 additional saliva samples collected from 40 - 65 year old men and women was determined using DRG SLV testosterone kit and the other method. From this study an R2 = 0.9866 was obtained with the following regression.
Image /page/1/Figure/3 description: This image is a scatter plot titled "DRG Comparison". The x-axis is labeled "LIA" and ranges from 0 to 700, while the y-axis is labeled "DRG ELISA" and also ranges from 0 to 700. The scatter plot shows a positive correlation between the two variables, and a line of best fit is drawn through the data points. The equation of the line of best fit is given as y = 1.0057x - 2.4196.
Sensitivity
The lowest analytical detectable level of testosterone that can be distinquished from the Zero Standard is 1.857 pg/mL at the 95 % confidence limit. The lowest functional sensitivity of 7.1 pg/mL at the 95% confidence limit was obtained.
Specificity
The following materials have been evaluated for cross reactivity. The percentage indicates cross reactivity at 50% displacement compared to Testosterone.
| Steroid | % Cross Reaction |
|---|---|
| Testosterone | 100% |
| 5α-Dihydrotestosterone | 0.80% |
| Androstenedione | 0.90% |
| 11β-hydroxysterone | 3.30% |
| 17α-methyltestosterone | 0.10% |
| 19-Nortestosterone | 3.30% |
| Epitestosterone | 0.10% |
| Estradiol | 0.10% |
| Progesterone | < 0,10% |
| Cortisol | < 0,10% |
| Estrone | < 0,10% |
| Danazol | < 0,10% |
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Reproducibility
Intra-Assay
The intra-assay variation was determined by 20 replicate measurements of 5 saliva samples within one run. The within-assay variability is shown below:
| Mean (pg/ml) | 144.00 | 256.15 | 81.30 | 35.35 | 12.94 |
|---|---|---|---|---|---|
| SD | 8.974 | 17.587 | 5.545 | 2.498 | 1.787 |
| CV (%) | 6.23 | 6.87 | 6.82 | 7.07 | 13.81 |
| n = | 20 | 20 | 20 | 20 | 20 |
Inter-Assay
The inter-assay (between-run) variation was determined by duplicate measurements of 5 saliva samples over 10 days.
| Mean (pg/mL) | 823.08 | 87.57 | 118.82 | 112.13 | 33.61 |
|---|---|---|---|---|---|
| SD | 45.314 | 6.864 | 8.864 | 8.628 | 3.232 |
| CV (%) | 5.51 | 7.84 | 7.46 | 7.69 | 9.62 |
| n = | 20 | 20 | 20 | 20 | 20 |
Inter-Lot
The Inter-Lot (between-lot) variation was determined by triplicate measurements of five saliva samples in three different kit lots. The between lot variability is shown below:
| Sample 1 | Sample 2 | Sample 3 | Sample 4 | Sample 5 | |
|---|---|---|---|---|---|
| Mean (pg/ml) | 64.5 | 352.89 | 517.65 | 44.00 | 116.54 |
| SD (pg/ml) | 3.77 | 13.44 | 15.01 | 1.53 | 5.00 |
| CV (%) | 5.85 | 3.81 | 2.90 | 3.47 | 4.29 |
| n = | 9 | 9 | 9 | 9 | 9 |
Recovery
Recovery of the DRG ELISA was determined by adding increasing amounts of the analyte to six different saliva samples containing different amounts of endogenous analyte. Each sample (native and spiked) was assayed and analyte concentrations of the samples were calculated from the standard curve. The percentage recoveries were determined by comparing expected and measured values of the samples
| Sample | Measured(pg/ml) | Expected(pg/ml) | Recovery(%) |
|---|---|---|---|
| 1 | 8.39 | - | - |
| 2396.90 | 2508.39 | 95.56 | |
| 517.17 | 508.39 | 101.73 | |
| 271.10 | 258.39 | 104.92 | |
| 55.95 | 58.39 | 95.82 | |
| 2 | 46.23 | - | - |
| 2474.70 | 2546.23 | 97.19 | |
| 564.37 | 546.23 | 103.32 | |
| 308.97 | 296.23 | 104.30 | |
| 88.87 | 96.23 | 92.35 | |
| 3 | 122.09 | - | - |
| 2602.41 | 2622.09 | 99.25 | |
| 591.16 | 622.09 | 95.03 |
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| 352.56 | 372.09 | 94.75 | |
|---|---|---|---|
| 166.40 | 172.09 | 96.69 | |
| 4 | 210.00 | - | - |
| 2650.00 | 2710.00 | 97.8 | |
| 700.00 | 710.00 | 98.6 | |
| 240.00 | 240.00 | 100.0 | |
| 5 | 1250.00 | - | - |
| 3700.00 | 3750.00 | 98.7 | |
| 1680.00 | 1750.00 | 96.0 | |
| 1250.00 | 1280.00 | 97.7 | |
| 6 | 2090.00 | - | - |
| 4550.00 | 4590.00 | 99.1 | |
| 2650.00 | 2590.00 | 102.3 | |
| 2130.00 | 2120.00 | 100.5 |
Linearity
Six saliva samples containing different amounts of analyte were serially diluted with zero standard and assayed with the DRG ELISA. Three native samples were serially diluted, and 3 samples were spiked with testosterone and then serially diluted up to 1:128. The percentage recovery was calculated by comparing the expected and measured values for testosterone. An assay linearity of 7.1 - 4500 pg/mL has been identified as the usable range. Samples above this range must be diluted and re-run.
| Sample 1 | Sample 2 | Sample 3 | Sample 4 | Sample 5 | Sample 6 | ||
|---|---|---|---|---|---|---|---|
| Concentration (pg/ml) | 4312.00 | 1838.00 | 440.00 | 8000.0 | 4500.0 | 5500.0 | |
| Average % Recovery | 97.5 | 99.6 | 98.9 | 100.3 | 100.8 | 100.7 | |
| Range of | from | 96.3 | 98.4 | 93.6 | 93.6 | 93.7 | 94.1 |
| Recovery % | to | 98.7 | 101.0 | 106.7 | 106.7 | 107.0 | 107.8 |
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DEPARTMENT OF HEALTH & HUMAN SERVICES
Image /page/4/Picture/1 description: The image shows the logo for the U.S. Department of Health and Human Services. The logo consists of a stylized caduceus, which is a symbol often associated with medicine and healthcare. The words "DEPARTMENT OF HEALTH & HUMAN SERVICES. USA" are arranged in a circular pattern around the caduceus.
Public Health Service
Food and Drug Administration 2098 Gaither Road Rockville MD 20850
JAN 2 7 2006
DRG International, Inc. c/o Mr. Gary Lehnus Lehnus & Associates 150 Cherry Land Rd. East Stroudsburg, PA 18301
Re: K052649
Trade/Device Name: DRG SLV Testosterone ELISA Test Regulation Number: 21 CFR§ 862.1680 Regulation Name: Testosterone test system Regulatory Class: Class I Product Code: CDZ Dated: December 27, 2005 Received: January 9, 2006
Dear: Mr. Lehnus
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.
If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to such additional controls. Existing major regulations affecting your device can be found in Title 21, Code of Federal Regulations (CFR), Parts 800 to 895. 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); and good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820).
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Page 2 -
This letter will allow you to begin marketing your device as described in your Section 510(k) premarket notification. The FDA finding of substantial equivalence of your device to a legally marketed predicate device results in a classification for your device and thus, permits your device to proceed to the market.
If you desire specific information about the application of labeling requirements to your device, or questions on the promotion and advertising of your device, please contact the Office of In Vitro Diagnostic Device Evaluation and Safety at (240) 276-0484. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR Part 807.97). You may obtain other general information on your responsibilities under the Act from the Division of Small Manufacturers, International and Consumer Assistance at its toll-free number (800) 638-2041 or (301) 443-6597 or at its Internet address http://www.fda.gov/cdrh/industry/support/index.html.
Sincerely vours.
Alberto Garcia
Alberto Gutierrez, Ph.D. Director Division of Chemistry and Toxicology Office of In Vitro Diagnostic Device Evaluation and Safety Center for Devices and Radiological Health
Enclosure
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Indications for Use
510(k) Number (if known): K052649
DRG SLV Testosterone ELISA Test Device Name:
Indications For Use:
An enzyme immunoassay for the in vitro diagnostic quantitative measurement of free active testosterone in saliva.
Measurement of testosterone is used in the diagnosis and treatment of disorders involving the male sex hormones (androgens), including primary and secondary hypogonadism, delayed or precocious puberty, impotence in males and, in females hirsutism (excessive hair) and virilization (masculinization) due to tumors, polycystic ovaries, and adrenogenital syndromes.
× Prescription Use (Part 21 CFR 801 Subpart D)
AND/OR
Over-The-Counter Use (21 CFR 801 Subpart C)
(PLEASE DO NOT WRITE BELOW THIS LINE-CONTINUE ON ANOTHER PAGE IF NEEDED)
Concurrence of CDRH, Office of Device Evaluation (OlVD)
Ann Chappie
Division Sign-Off
Office of In Vitro Diagnostic Device Evaluation and Safety
710(k) K052649
Page of 1
§ 862.1680 Testosterone test system.
(a)
Identification. A testosterone test system is a device intended to measure testosterone (a male sex hormone) in serum, plasma, and urine. Measurement of testosterone are used in the diagnosis and treatment of disorders involving the male sex hormones (androgens), including primary and secondary hypogonadism, delayed or precocious puberty, impotence in males and, in females hirsutism (excessive hair) and virilization (masculinization) due to tumors, polycystic ovaries, and adrenogenital syndromes.(b)
Classification. Class I.