(245 days)
Not Found
No
The summary describes a standard enzymatic assay performed on an automated chemistry analyzer. There is no mention of AI, ML, or any computational methods beyond basic data processing for calculating results and controlling the instrument.
No
This device is an in-vitro diagnostic assay used for the quantitative determination of HbA1c in venous whole blood samples for monitoring long-term glucose control in persons with diabetes. It is not used for direct treatment or diagnosis, but rather provides information for clinical decision-making.
No
Explanation: The "Intended Use / Indications for Use" section explicitly states, "This test is not to be used to diagnose or screen for diabetes." Instead, it is used for "monitoring long-term glucose control of persons with diabetes."
No
The device is a test kit comprised of reagents, calibrators, and controls, which are physical components. While it is used with an automated chemistry analyzer that has software, the device itself is not solely software.
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 test kit is intended for "in-vitro diagnostic use only." It also describes the quantitative determination of stable HbA in venous whole blood samples, which is a diagnostic measurement performed outside of the body.
- Device Description: The description details reagents, calibrators, and controls used for testing biological samples (whole blood).
- Performance Studies: The document describes various performance studies (precision, linearity, method comparison) conducted to validate the assay's performance for diagnostic purposes in a clinical laboratory setting.
- Intended User / Care Setting: The intended user is a "clinical laboratory," which is where IVD tests are typically performed.
All of these points align with the definition of an In Vitro Diagnostic device.
N/A
Intended Use / Indications for Use
Diazyme Direct HbA1c Assay (Enzymatic, On-Board Lysis) test kit is intended for use in the quantitative determination of stable HbA for use in monitoring long-term glucose control of persons with diabetes. For in-vitro diagnostic use only.
Diazyme Direct HbA1c Assay Calibrator Set is intended to be used for calibration of Diazyme Direct HbA1c Assay (Enzymatic, On-Board Lysis). For in-vitro diagnostic use only.
Diazyme Direct HbA1c Assay Control Set is intended to be used for quality control by monitoring accuracy and precision of Diazyme Direct HbA1c Assay (Enzymatic, On-Board Lysis). For in-vitro diagnostic use only.
Product codes (comma separated list FDA assigned to the subject device)
LCP, JIT, JJX
Device Description
The Diazyme Direct HbA1c Assay (Enzymatic, On-Board Lysis) test kit is comprised of Reagent 1, Reagent 2, and Lysis Buffer. The Diazyme Direct HbA1c Assay Calibrator Set contains 2 levels of human whole blood based lyophilized calibrators. The Diazyme Direct HbA1c Assay Control Set contains 2 levels of human whole blood based lyophilized controls. The device performs an enzymatic assay where lysed whole blood samples undergo protease digestion, releasing glycated valines. These glycated valines act as substrates for a specific recombinant fructosyl valine oxidase (FVO) enzyme, which cleaves N-terminal valines and produces hydrogen peroxide. The hydrogen peroxide is then measured using a horseradish peroxidase (POD) catalyzed reaction and a suitable chromogen. This direct assay does not require a separate measurement for total Hemoglobin (Hb). The test is intended for quantitative determination of stable HbA1c in venous whole blood samples with on-board blood lysis. The device is used with an automated chemistry analyzer (Roche Modular P), which performs automatic on-board 1:12.5 dilution of the sample with lysis buffer.
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
Clinical laboratory
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
Precision Summary - Internal Precision:
- Sample Size: 5 unaltered whole blood specimens (4.6%, 5.4%, 7.5%, 9.7%, and 11.9% HbA1c), 3 lots of HbA1c calibrators (2 levels each), and 3 lots of HbA1c controls (2 levels each).
- Data Source: Whole blood samples were IRB approved.
- Annotation Protocol: Tested according to CLSI EP5-A2 guideline. All samples were tested 2 runs per day, 2 replicates per run over 20 working days. Data analyzed with EP Evaluator (Release 11).
Precision Summary - Multiple Site Precision Study:
- Sample Size: 5 whole blood samples (4.6%, 5.4%, 7.5%, 9.7%, and 11.9% HbA1c), 3 lots of calibrators, and 3 lots of controls.
- Data Source: Same sets of samples used at all sites.
- Annotation Protocol: Evaluated according to a modified CLSI EP5-A2 guideline. Samples tested in duplicates per run, 2 runs per day for 5 working days with the same lot of the reagent. Testing conducted at 3 different sites, by 3 different operators, on 3 different Modular P instruments.
Method Comparison Study:
- Sample Size: Total of 124 unique samples at site 1, total of 132 unique samples at site 2, and a total of 120 unique samples at site 3.
- Data Source: Individual K2 EDTA whole blood samples (IRB approved).
- Annotation Protocol: Followed CLSI: EP9-A2 guideline. Samples tested using the same lot of the reagents at three different sites on three different Modular P analyzers by three different operators, in comparison with Tosoh G8 HPLC HbA1c method (NGSP certification method).
Analytical specificity (Endogenous substances):
- Sample Size: Three whole blood samples with "low", "medium", and "high" HbA1c concentrations.
- Data Source: Whole blood samples.
- Annotation Protocol: Followed CLSI EP7-A2. Each whole blood sample was spiked with potential endogenous interference substances per protocol and tested in triplicates on the Roche Modular P analyzer.
Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)
Precision Summary - Internal Precision:
- Study Type: Internal Precision
- Sample Size: 240 replicates per sample/control/calibrator across 20 working days, 3 reagent lots.
- Key Results:
- Whole blood samples spanning AMR (4.6% to 11.9% HbA1c): Total CV% ranged from 0.9% to 1.7%.
- Controls: Total CV% ranged from 0.7% to 1.2%.
- Calibrators: Total CV% ranged from 0.7% to 1.2%.
- Conclusion: Within-run, between-run, between-day, between-lot and total CV% were less than 2%, meeting acceptance criteria.
Precision Summary - Multiple Site Precision Study:
- Study Type: Multiple Site Precision Study
- Sample Size: 60 replicates per sample/control/calibrator across 5 working days at 3 sites.
- Key Results:
- Whole blood samples: Total CV% ranged from 1.2% to 1.4%. Between-Site CV% ranged from 0.9% to 1.4%.
- Controls: Total CV% ranged from 0.6% to 1.0%. Between-Site CV% ranged from 0.5% to 1.0%.
- Calibrators: Total CV% ranged from 0.7% to 1.1%. Between-Site CV% ranged from 0.8% to 1.0%.
- Conclusion: All samples tested showed that the within-run precision, between-run, between-day, between-site, and total CV% were less than 2% meeting the acceptance criteria.
LOD/LOB/LOQ:
- Study Type: Limit of Blank (LOB), Limit of Detection (LOD), Limit of Quantitation (LOQ) determination.
- Key Results:
- LOB: 0.2% HbA1c (maximal value among 3 reagent lots).
- LOD: 0.5% HbA1c (maximal value among 3 reagent lots).
- LOQ: 0.8% HbA1c (for all 3 reagent lots).
Linearity:
- Study Type: Linearity
- Sample Size: 11 levels of whole blood linearity set.
- Key Results: Bias ≤10%. The assay is linear from 4% to 12% HbA1c.
Method Comparison Study:
- Study Type: Method Comparison (comparison to Tosoh G8 HPLC HbA1c method).
- Sample Size: 124 unique samples (site 1), 132 unique samples (site 2), 120 unique samples (site 3).
- Key Results (Regular Linear Regression):
- Combined Slope: 1.023 (95% CI: 1.011 - 1.035)
- Combined Intercept: -0.090 (95% CI: -0.180 to 0.000)
- Combined Correlation Coefficient(R): 0.9937
- Sample Range (Diazyme): 4.2 - 12.0
- Key Results (Deming Regression):
- Combined Slope: 1.030 (95% CI: 1.018 – 1.042)
- Combined Intercept: -0.140 (95% CI: -0.230 to -0.050)
- Combined Correlation Coefficient(R): 0.9937
- Sample Range (Diazyme): 4.2 - 12.0
- Conclusion: Method comparison data analysis demonstrated that the results met the acceptance criteria.
Analytical specificity:
- Study Type: Interference study for endogenous substances.
- Key Results: No significant interference (10%) may result in inaccurate HbA1c values.
Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)
Precision (Total CV%):
- Internal Precision: Samples 0.9% - 1.7%, Controls 0.7% - 1.2%, Calibrators 0.7% - 1.2%.
- Multiple Site Precision: Samples 1.2% - 1.4%, Controls 0.6% - 1.0%, Calibrators 0.7% - 1.1%.
Correlation Coefficient (R): 0.9937 (Combined, Regular Linear and Deming)
Linearity Bias: ≤10%
Interference (Deviation):
§ 864.7470 Glycosylated hemoglobin assay.
(a)
Identification. A glycosylated hemoglobin assay is a device used to measure the glycosylated hemoglobins (A1a , A1b , and A1c ) in a patient's blood by a column chromatographic procedure. Measurement of glycosylated hemoglobin is used to assess the level of control of a patient's diabetes and to determine the proper insulin dosage for a patient. Elevated levels of glycosylated hemoglobin indicate uncontrolled diabetes in a patient.(b)
Classification. Class II (performance standards).
0
Image /page/0/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo features a stylized depiction of a human figure, represented by three overlapping profiles, with the head tilted upwards. The profiles are rendered in a dark color, creating a sense of depth and unity. The text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" is arranged in a circular pattern around the figure.
Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002
November 21, 2016
DIAZYME LABORATORIES ABHIJIT DATTA DIRECTOR, TECHNICAL OPERATIONS 12889 GREGG COURT POWAY CA 92064
Re: K160762
Trade/Device Name: Diazyme Direct HbA1c Assay (Enzymatic, On-Board Lysis), Diazyme Direct HbA1c Assay Calibrator Set, Diazyme Direct HbA1c Assay Control Set Regulation Number: 21 CFR 864.7470 Regulation Name: Glycosylated hemoglobin assay Regulatory Class: II Product Code: LCP, JIT, JJX Dated: October 5, 2016 Received: October 6, 2016
Dear Abhijit Datta:
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,
Katherine Serrano -S
For : 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) K160762
Device Name
Diazyme Direct HbA1c Assay (Enzymatic, On-Board Lysis), Diazyme Direct HbA1c Assay Calbrator Set, Diazyme Direct HbA1c Assay Control Set
Indications for Use (Describe)
Diazyme Direct HbA 1c Assay (Enzymatic, On-Board Lysis) test kit is intended for use in the quantitative determination of stable HbA in venous whole blood samples with on-board blood lysis application in a clinical laboratory. This test is not to be used to diagnose or screen for diabetes. The measurement of HbA 1c concentration is for use in monitoring longterm glucose control of persons with diabetes. For in-vitro diagnostic use only.
Diazyme Direct HbA1c Assay Calibrator Set is intended to be used for calibration of Diazyme Direct HbA1c Assay (Enzymatic, On-Board Lysis). For in-vitro diagnostic use only.
Diazyme Direct HbA1c Assay Control Set is intended to be used for quality control by monitoring accuracy and precision of Diazyme Direct HbA1c Assay (Enzymatic, On-Board Lysis). For in-vitro diagnostic use only.
| Type of Use (Select one or both, as applicable) | |
|---|---|
| ------------------------------------------------- | -- |
X Prescription Use (Part 21 CFR 801 Subpart D)
Over-The-Counter Use (21 CFR 801 Subpart C)
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K160762
510k Summary
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.
| Submitter's name: | Diazyme Laboratories |
|---|---|
| Submitter's address: | 12889 Gregg Court |
| Poway, CA 92064 | |
| USA | |
| Name of Contact Person: | Dr. Abhijit Datta |
| Diazyme Laboratories | |
| 12889 Gregg Court | |
| Poway, CA 92064 | |
| Phone: 858-455-4762 | |
| Fax: 858-455-2120 | |
| Date the Summary was Prepared: | November 15, 2016 |
| Name of the Device | Diazyme Direct HbA1c Assay (Enzymatic, On-Board Lysis) |
| Diazyme Direct HbA1c Assay Calibrator Set | |
| Diazyme Direct HbA1c Assay Control Set | |
| Trade Name: | Diazyme Direct HbA1c Assay (Enzymatic, On-Board Lysis) |
| Diazyme Direct HbA1c Assay Calibrator Set | |
| Diazyme Direct HbA1c Assay Control Set | |
| Common Name: | Direct Enzymatic Assay, HbA1c |
| Classification Name: | Glycosylated Hemoglobin Assay |
Establishment Registration
The Device Establishment Registration Number is 2032900.
4
Regulatory Information:
| Regulation Description | Product Code | Device Class | Regulation | Panel |
|---|---|---|---|---|
| Glycosylated | ||||
| Hemoglobin Assay | LCP | II | 21 CFR 864.7470 | Hematology, 81 |
| Calibrator | JIT | II | 21 CFR §862.1150 | Chemistry, 75 |
| Quality Control | ||||
| Material | JJX | I | 21 CFR §862.1660 | Chemistry, 75 |
Submission type: Traditional 510k
Predicate Device
Diazyme Laboratories claims substantial equivalence to the currently marketed Diazyme Direct HbA1c Enzymatic Assay. The reference numbers are K070743.
Performance Standards
At this time, no special controls under Section 513 or performance standard under Section 514 have been issued for in vitro diagnostic products.
Manufacturing Address
The Diazyme Direct HbA1c Assay (Enzymatic, On-Board Lysis) will be manufactured and distributed by Diazyme Laboratories.
Manufacturing Site Address Diazyme Laboratories 12889 Gregg Court Poway, CA 92064
Description of the Device
Clinical Significance
Hemoglobin A1c is an important test recommended by the American Diabetes Association (ADA) and its usefulness was clarified by the United Kingdom Prospective Diabetes Study (UKPDS) and Diabetes Control and Complications Trial (DCCT). Currently, as part of the diabetes management program, HbA1c testing should be performed at least biannually in all patients and quarterly for patients whose therapy has changed or who are not meeting treatment goals. Glycohemoglobin is produced by non-enzymatic addition of glucose to amino groups in hemoglobin. HbA1c refers to glucose modified hemoglobin A (HbA) specifically at N-terminal
5
valine residues of hemoglobin beta chains. HbA1c test is used both as an index of mean glycemia and as a measure of risk for the development of diabetes complications. Therefore, the HbA1c test is a good indicator of glycemic control in the preceding 2-3 months.
Assay Principle
Diazyme Direct HbA1c test is an enzymatic assay in which lysed whole blood samples are subjected to extensive protease digestion with Bacillus sp protease. This process releases amino acids including glycated valines from the hemoglobin beta chains. Glycated valines then serve as substrates for specific recombinant fructosyl valine oxidase (FVO) enzyme. The recombinant FVO specifically cleaves N-terminal valines and produces hydrogen peroxide. This, in turn, is measured using a horseradish peroxidase (POD) catalyzed reaction and a suitable chromogen. No separate measurement for total Hemoglobin (Hb) is needed in this Direct HbA1c Assay.
Detailer Device Description
-
- The Diazyme Direct HbA1c assay kit (Enzymatic, On-Board Lysis) is comprised of a Reagent 1, Reagent 2, Lysis Buffer.
-
- Diazyme Direct HbA1c Assay Calibrator Set: 2 levels, human whole blood based in lyophilized form. It is intended to be used for calibration of Diazyme Direct HbA1c Assay (Enzymatic, On-Board Lysis).
-
- Diazyme Direct HbA1c Assay Control Set: 2 levels, human whole blood based in lyophilized form. It is intended to be used for quality control by monitoring accuracy and precision of Diazyme Direct HbA1c Assay (Enzymatic, On-Board Lysis).
The verification and/or validation studies were performed on the automated chemistry analyzer Roche Modular P (K953239/005). Modular P analyzer is a high through-put automated chemistry analyzer and that can process 800 samples per hour. The predicate performance studies at submission were performed on Hitachi 917 which is an obsolete member of the Roche Mod P Roche family of instruments. The Control Unit of the Roche Modular P analyzer uses a graphical interface to control all instrument functions. The computer, keyboard, and touchscreen monitor allows users to navigate through the software, enter assay, calibrator, and control information, and make test selections. The Diazyme Direct HbA1c Assay Modular P application parameters provided are programmed into the Modular P analyzers. The reagents, calibrators and controls are loaded into the analyzer. The Roche Modular P stores the Diazyme Direct HbA1c Assay reagents in a refrigerated compartment. Reagent and sample pipettes automatically aspirate and dispense specified amounts of reagents or calibrators, controls, and samples into reaction cells. The change in absorbance is measured at specified wavelengths.
Assay Scheme for Roche Modular P Analyzer
Use the following scheme as a guideline for analyzer application. Note: Diazyme Direct HbA1c Assay is an end-point assay and the first reading point A1 is right before the addition of reagent R2.
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Image /page/6/Figure/0 description: This image shows a timeline of a process. At 0 minutes, 10 uL of blood, 125 uL of lysis buffer, and 128 uL of R1 are added. At 5 minutes, 56 uL of R2 is added, and A1 is measured at 700 nm. At 8 minutes, A2 is measured.
After a 2-point calibration, linear-fitting is used to fit calibration curve through mean values of the response for calibrators of known concentrations. The Roche Modular P calculates the HbA1c% concentration of a patient sample by interpolation of the obtained signal to a stored 2point calibration curve.
#: The Modular P analyzer performs automatic on-board 1: 12.5 dilution using lysis buffer (10 uL sample + 125 uL lysis buffer) for rapid blood sample lysis.
The following key parameters are input into the Mod P user defined channel interface using the provided Diazyme Direct HbA1c Assay (DZ168C) Parameter sheet.
| Modular P Applications | |
|---|---|
| Measuring mode: | Absorbance |
| Abs. calculation mode: | 2 Point END |
| Reaction mode: | S-R1/R3 |
| Reaction direction: | Increase |
| Primary/Secondary Wavelength: | 700nm/800nm |
| Calc. first/last: | 16/28 |
| Unit: | % |
| Pipetting parameters | |
| R1: | 128 μL |
| Sample: | 10 µL; Sample Diluent (lysis buffer) 125 µL (1:12.5 dilution) |
| Diluted Sample: | 20 μL |
| R2: | 56 µL |
| Total volume: | 204 µL |
The Design control verification and/or validation studies were performed on Roche Mod P automated analyzer.
Intended Use of the Device:
Diazyme Direct HbA1c Assay (Enzymatic, On-Board Lysis) test kit is intended for use in the quantitative determination of stable HbA1c in venous whole blood samples with on-board blood lysis application in a clinical laboratory. This test is not to be used to diagnose or screen for
7
diabetes. The measurement of HbA1c concentration is for use in monitoring long-term glucose control of persons with diabetes. For in-vitro diagnostic use only.
Diazyme Direct HbA1c Assay Calibrator Set is intended to be used for calibration of Diazyme Direct HbA1c Assay (Enzymatic, On-Board Lysis). For in-vitro diagnostic use only.
Diazyme Direct HbA1c Assay Control Set is intended to be used for quality control by monitoring accuracy and precision of Diazyme Direct HbA1c Assay (Enzymatic, On-Board Lysis). For in-vitro diagnostic use only.
Performance Characteristics of Device
Precision Summary
- Internal Precision
Internal precision of the Diazyme Direct HbA1c Assay (Enzymatic, On-Board Lysis) was evaluated according to CLSI EP5-A2 guideline. This assessment was performed on the Roche Modular P analyzer at Diazyme Laboratories.
In the study, three lots of reagents were used. For each lot of reagent, 5 unaltered whole blood specimens containing 4.6%, 5.4%, 7.5%, 9.7% and 11.9% HbA1c spanning AMR were tested. Three lots of HbA1c calibrators and three lots of HbA1c controls, two levels each, were tested as samples to evaluate precision. All samples were tested 2 runs per day, 2 replicates per run over 20 working days according to CLSI EP5-A2 protocol. Whole blood samples used in the study were IRB approved.
Precision data for the three lots of reagents tested are given below. Data analyzed with EP Evaluator (Release 11).
| Sample | Mean
(N=240) | | Within-
Run | Between-
Run | | Between-
day | | Between
lot | | Total | |
|-------------|-----------------|------|----------------|-----------------|------|-----------------|------|----------------|------|-------|------|
| | | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| Sample 1 | 4.64 | 0.04 | 0.8% | 0.07 | 1.5% | 0.00 | 0.0% | 0.08 | 1.6% | 0.08 | 1.7% |
| Sample 2 | 5.36 | 0.05 | 0.9% | 0.05 | 0.9% | 0.00 | 0.0% | 0.07 | 1.2% | 0.07 | 1.2% |
| Sample 3 | 7.51 | 0.05 | 0.6% | 0.05 | 0.7% | 0.00 | 0.0% | 0.07 | 0.9% | 0.07 | 0.9% |
| Sample 4 | 9.61 | 0.06 | 0.6% | 0.05 | 0.5% | 0.03 | 0.3% | 0.08 | 0.9% | 0.08 | 0.9% |
| Sample 5 | 11.89 | 0.09 | 0.7% | 0.08 | 0.6% | 0.04 | 0.4% | 0.12 | 1.0% | 0.12 | 1.0% |
| Con 1 Lot 1 | 6.22 | 0.05 | 0.8% | 0.03 | 0.5% | 0.01 | 0.1% | 0.06 | 1.0% | 0.06 | 1.0% |
| Con 2 Lot 1 | 9.47 | 0.06 | 0.6% | 0.04 | 0.4% | 0.02 | 0.2% | 0.07 | 0.8% | 0.07 | 0.8% |
| Con 1 Lot 2 | 5.70 | 0.04 | 0.8% | 0.04 | 0.7% | 0.02 | 0.4% | 0.06 | 1.1% | 0.06 | 1.1% |
| Con 2 Lot 2 | 9.11 | 0.05 | 0.5% | 0.04 | 0.4% | 0.03 | 0.3% | 0.07 | 0.7% | 0.07 | 0.7% |
| Con 1 Lot 3 | 6.04 | 0.05 | 0.7% | 0.05 | 0.9% | 0.00 | 0.0% | 0.07 | 1.1% | 0.07 | 1.2% |
| Con 2 Lot 3 | 9.68 | 0.05 | 0.6% | 0.04 | 0.4% | 0.00 | 0.0% | 0.07 | 0.7% | 0.07 | 0.7% |
| Cal 1 Lot 1 | 6.22 | 0.04 | 0.6% | 0.04 | 0.6% | 0.00 | 0.0% | 0.05 | 0.8% | 0.05 | 0.9% |
| Cal 2 Lot 1 | 12.30 | 0.07 | 0.6% | 0.04 | 0.3% | 0.03 | 0.2% | 0.09 | 0.7% | 0.09 | 0.7% |
| Cal 1 Lot 2 | 5.68 | 0.05 | 0.9% | 0.04 | 0.7% | 0.02 | 0.4% | 0.07 | 1.1% | 0.07 | 1.2% |
| Cal 2 Lot 2 | 9.70 | 0.06 | 0.6% | 0.04 | 0.4% | 0.02 | 0.2% | 0.07 | 0.8% | 0.07 | 0.8% |
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| Cal 1 Lot 3 | 6.04 | 0.05 | 0.8% | 0.04 | 0.7% | 0.00 | 0.0% | 0.06 | 1.0% | 0.06 | 1.1% |
|---|---|---|---|---|---|---|---|---|---|---|---|
| Cal 2 Lot 3 | 11.30 | 0.06 | 0.5% | 0.04 | 0.3% | 0.01 | 0.1% | 0.07 | 0.6% | 0.07 | 0.6% |
Conclusion: 20-day reproducibility data for all samples tested showed that the within-run, between-run, between-day, between-lot and total CV% were less than 2%, meeting acceptance criteria.
- Multiple Site Precision Study (Three sites)
Multiple site precision study was performed at two external sites and the Diazyme site. In this study, same sets of samples were used at all sites. Multiple site precision of the Diazyme Direct HbA1c Assay was evaluated according to a modified CLSI EP5-A2 guideline on Roche Modular P analyzer. Five whole blood samples containing 4.6%, 5.4%, 7.5%, 9.7% and 11.9% HbA1c spanning assay AMR, three lots of calibrators and three lots of controls were tested in duplicates per run, 2 runs per day for 5 working days with the same lot of the reagent to evaluate between site assay precision. Testing was conducted at 3 different sites, by 3 different operators, on 3 different Modular P instruments. Within-run, between-day, between-site, and total precision were calculated.
| Sample | Mean | Within-Run | Between-Run | Between-day | Total | ||||
|---|---|---|---|---|---|---|---|---|---|
| (n=60) | SD | %CV | SD | %CV | SD | %CV | SD | %CV | |
| Sample 1 | 4.67 | 0.05 | 1.0% | 0.04 | 0.8% | 0.02 | 0.5% | 0.07 | 1.4% |
| Sample 2 | 5.37 | 0.04 | 0.8% | 0.05 | 1.0% | 0.00 | 0.0% | 0.07 | 1.2% |
| Sample 3 | 7.52 | 0.05 | 0.7% | 0.06 | 0.8% | 0.00 | 0.0% | 0.08 | 1.0% |
| Sample 4 | 9.67 | 0.07 | 0.8% | 0.11 | 1.1% | 0.00 | 0.0% | 0.13 | 1.4% |
| Sample 5 | 11.92 | 0.09 | 0.8% | 0.09 | 0.8% | 0.06 | 0.5% | 0.14 | 1.2% |
| Con1 Lot 1 | 6.21 | 0.03 | 0.6% | 0.00 | 0.0% | 0.00 | 0.0% | 0.03 | 0.6% |
| Con2 Lot 1 | 9.48 | 0.05 | 0.6% | 0.05 | 0.6% | 0.03 | 0.3% | 0.08 | 0.9% |
| Con1 Lot 2 | 5.63 | 0.04 | 0.7% | 0.02 | 0.4% | 0.03 | 0.6% | 0.06 | 1.0% |
| Con2 Lot 2 | 9.11 | 0.05 | 0.5% | 0.04 | 0.4% | 0.03 | 0.3% | 0.07 | 0.7% |
| Con1 Lot 3 | 6.01 | 0.05 | 0.8% | 0.01 | 0.2% | 0.02 | 0.3% | 0.06 | 0.9% |
| Con2 Lot 3 | 9.65 | 0.06 | 0.7% | 0.05 | 0.5% | 0.04 | 0.4% | 0.09 | 1.0% |
| Call Lot 1 | 6.21 | 0.04 | 0.7% | 0.03 | 0.5% | 0.00 | 0.0% | 0.06 | 0.9% |
| Cal2 Lot 1 | 12.32 | 0.07 | 0.6% | 0.05 | 0.4% | 0.05 | 0.4% | 0.10 | 0.8% |
| Call Lot 2 | 5.63 | 0.06 | 1.0% | 0.00 | 0.0% | 0.02 | 0.3% | 0.06 | 1.1% |
| Cal2 Lot 2 | 9.74 | 0.06 | 0.6% | 0.06 | 0.7% | 0.03 | 0.4% | 0.10 | 1.0% |
| Call Lot 3 | 6.02 | 0.05 | 0.8% | 0.03 | 0.4% | 0.01 | 0.1% | 0.05 | 0.9% |
| Cal2 Lot 3 | 11.36 | 0.06 | 0.6% | 0.08 | 0.7% | 0.05 | 0.5% | 0.12 | 1.0% |
Multiple site precision data is summarized in the following table:
| Mean | Between-Site | ||
|---|---|---|---|
| Sample | (n=60) | SD | %CV |
| Sample 1 | 4.7 | 0.07 | 1.4 |
| Sample 2 | 5.4 | 0.06 | 1.2 |
| Sample 3 | 7.5 | 0.07 | 0.9 |
| Sample 4 | 9.7 | 0.12 | 1.3 |
9
| Sample 5 | 11.9 | 0.14 | 1.2 |
|---|---|---|---|
| Control 1 Lot 1 | 6.2 | 0.03 | 0.5 |
| Control 2 Lot 1 | 9.5 | 0.08 | 0.9 |
| Control 1 Lot 2 | 5.6 | 0.06 | 1.0 |
| Control 2 Lot 2 | 9.1 | 0.07 | 0.7 |
| Control 1 Lot 3 | 6.0 | 0.06 | 0.9 |
| Control 2 Lot 3 | 9.6 | 0.09 | 0.9 |
| Calibrator 1 Lot 1 | 6.2 | 0.05 | 0.8 |
| Calibrator 2 Lot 1 | 12.3 | 0.10 | 0.8 |
| Calibrator 1 Lot 2 | 5.6 | 0.05 | 1.0 |
| Calibrator 2 Lot 2 | 9.7 | 0.10 | 1.0 |
| Calibrator 1 Lot 3 | 6.0 | 0.05 | 0.9 |
| Calibrator 2 Lot 3 | 11.4 | 0.12 | 1.0 |
Conclusion: All samples tested showed that the within-run precision, between-run, between-day, between-site, and total CV% were less than 2% meeting the acceptance criteria.
LOD/LOB/LOQ
The LOB, LOD, and LOQ of the Diazyme Direct HbA1c Assay (Enzymatic, On-Board Lysis Assay were determined according to CLSI EP17-A2 with three lots of the reagents.
The LOB was determined as 0.0%, 0.1% and 0.2% for reagent lot 1, lot 2, and lot 3 respectively. The HbA1c Assay LOB is obtained as the maximal value between all three reagents lots, 0.2%.
The LOD was determined as 0.38%. 0.43%, and 0.5% for reagent lot 1. lot 2. and lot 3 respectively. The Direct HbA1c Assay LOD is obtained as the maximal value among the three lots, 0.5%.
Using the calculated CV's from the 5 concentrations, curve fit analysis was performed to determine the upper 95% confidence level of point with CV less than 20%. EP Evaluator (Release 11) analysis showed LOQ = 0.8% for Reagent Lots 1, 2, and 3, respectively.
Linearity
11 levels of whole blood linearity set were prepared by dilution of two whole blood samples, Low and high samples with known HbA1c of 3.8% and 12.3% respectively were used to generate the test levels according to CLSI EP6-A and tested in triplicate, again, per guideline. Data analysis showed bias ≤10%. The assay is linear from 4% to12% HbA1c.
Calibrator Traceability
Diazyme Direct HbA1c Assay is traceable to NGSP/DCCT. The assay methodology is traceable to IFCC per NGSP correlation guideline (www.ngsp.org).
Method Comparison Study
10
Following the CLSI: EP9-A2 guideline, the Diazyme Direct HbA1c Assay (Enzymatic, On-Board Lysis) was evaluated on the Roche Modular P testing individual K2 EDTA whole blood samples (IRB approved) in comparison with Tosoh G8 HPLC HbA1c method, which is used as the NGSP certification method in NGSP reference laboratories (www.NGSP.org). A total of 124 unique samples at site 1, total of 132 unique samples at site 2, and a total of 120 unique samples at site 3 were tested using the same lot of the reagents at three different sites on three different Modular P analyzers by three different operators.
Summary of Regression Analysis: Regular Linear
| Parameter | Site 1 | Site 2 | Site 3 | Combined |
|---|---|---|---|---|
| Slope | 1.006 | 1.033 | 1.026 | 1.023 |
| 95% CI | 0.983 - 1.030 | 1.015 - 1.051 | 1.005 - 1.047 | 1.011 - 1.035 |
| Intercept | 0.04 | -0.126 | -0.18 | -0.090 |
| 95% CI | -0.13 to 0.21 | -0.268 to 0.015 | -0.34 to -0.01 | -0.180 to 0.000 |
| Correlation | ||||
| Coefficient(R) | 0.9918 | 0.9950 | 0.9938 | 0.9937 |
| Sample Range | ||||
| (Diazyme) | 4.4 - 12.0 | 4.2 - 11.9 | 4.4 - 12.0 | 4.2 - 12.0 |
Summary of Regression Analysis: Deming
| Parameter | Site 1 | Site 2 | Site 3 | Combined |
|---|---|---|---|---|
| Slope | 1.015 | 1.039 | 1.033 | 1.030 |
| 95% CI | 0.992 – 1.038 | 1.021 - 1.057 | 1.012 - 1.054 | 1.018 – 1.042 |
| Intercept | -0.02 | -0.167 | -0.23 | -0.140 |
| 95% CI | -0.19 to 0.15 | -0.309 to -0.026 | -0.40 to -0.06 | -0.230 to -0.050 |
| Correlation | ||||
| Coefficient(R) | 0.9918 | 0.9950 | 0.9938 | 0.9937 |
| Sample Range | ||||
| (Diazyme) | 4.4 - 12.0 | 4.2 - 11.9 | 4.4 - 12.0 | 4.2 - 12.0 |
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Conclusion: Method comparison data analysis of the Diazyme Direct HbA1c Assay (Enzymatic, On-Board Lysis) versus the predicate Tosoh HPLC Assay testing K2 EDTA whole blood at three different sites on three different Modular P analyzers by three different operators demonstrated the results met the acceptance criteria.
Analytical specificity
Endogenous substances:
To determine the level of interference from substances present whole blood samples, the Diazyme Direct HbA1c Assay (Enzymatic, On-Board Lysis) was used to test three whole blood samples, which contained "low", "medium", and "high" HbA1c concentrations, following CLSI EP7-A2. To evaluate interference, each whole blood samples were spiked with potential endogenous interference substances per protocol and tested in triplicates on the Roche Modular P analyzer.
| Interfering Substances | Concentration |
|---|---|
| Ascorbic Acid | 12 mg/dL |
| Bilirubin | 15 mg/dL |
| Bilirubin Conjugated | 13 mg/dL |
| Hemoglobin | 21 mg/dL |
| Triglycerides | 4000 mg/dL |
| Glucose | 4000 mg/dL |
| Uric Acid | 30 mg/dL |
| Urea | 80 mg/dL |
| Acetaminophen | 20 mg/dL |
| Acetylsalicylic Acid | 65.2 mg/dL |
| Metformin | 4 mg/dL |
| Ibuprofen | 50 mg/dL |
| Glyburide | 0.19 mg/dL |
| Total Protein | 21000 mg/dL |
| Vitamin E | 13.6 mg/dL |
| Rheumatoid Factor | 375 IU/mL |
The following substances showed no significant interference (10%) may result in inaccurate HbA1c values.
12
Substantial Equivalence
Device Modifications
The intended use of the modified device and fundamental scientific technology used in the test has not changed as a result of the modification.
Specific modifications:
-
- Change in number of reagents supplied with on-board lysis application
-
- Change in concentration of reagent components
-
- Change in product labeling to reflect the modification
Summary of Assay Kit Components
| Predicate device K070734 | Subject Device |
|---|---|
| Lysis buffer | Lysis buffer |
| 100mM CHES pH 8.7, 1% Triton X 100, 0.45% SDS, 0.5mM redox agent | >100mM Tris pH >8.0, 1% Triton X 100, |
1.5% nonionic and ionic detergents,
4KU/mL proteases |
| Reagent1a | Reagent 1 |
| 5mM MES pH 7.0, >4KU/mL proteases,
10µM redox agent | 5mM MES pH >6.0, 10U/mL FVO enzyme,
90U/mL POD, 0.8mM chromagen | >5mM bis-Tris pH>7.0, >10U/mL FVO
enzyme, 90U/mL POD, >50μΜ chromagen |
| None | None |
| HbA1c Calibrator set
(linear mode, lyophilized whole blood based) | HbA1c Calibrator set
(linear mode, lyophilized whole blood based) |
| 1 x 0.5mL Level 1 | 1 x 0.5mL Level 1 |
| 1 x 0.5mL Level 2 | 1 x 0.5mL Level 2 |
| HbA1c Control set
(lyophilized whole blood based) | HbA1c Control set
(lyophilized whole blood based) |
| 1 x 0.5mL Level 1 | 1 x 0.5mL Level 1 |
Substantial Equivalence Comparison Table
Indications for Use
| Direct HbA1c Enzymatic Assay
| (K070734) | Direct HbA1c Assay (Enzymatic, On-Board Lysis) (modified device; special 510k submission) | Equivalency |
|---|---|---|
| Diazyme Direct Enzymatic Hemoglobin | Diazyme Direct HbA1c Assay | Same |
13
| A1c (glycated hemoglobin A1c; A1c;
HbA1c) reagents are intended for use in
the quantitative determination of stable
HbA1c in human whole blood samples.
Measurement of hemoglobin A1c is a
valuable indicator for long-term diabetic
control. For in-vitro diagnostic use only. | (Enzymatic, On-Board Lysis) test kit is
intended for use in the quantitative
determination of stable HbA1c in venous
whole blood samples with on-board
blood lysis application in a clinical
laboratory. This test is not to be used to
diagnose or screen for diabetes. The
measurement of HbA1c concentration is
for use in monitoring long-term glucose
control of persons with diabetes. For in-
vitro diagnostic use only. |
| --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- | --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- |
|---|
Principle
| Direct HbA1c Enzymatic Assay
(K070734) | Direct HbA1c Assay (Enzymatic, On-
Board Lysis) | Equivalency |
|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-------------|
| Direct Enzymatic HbA1c test is an
enzymatic assay in which lysed whole
blood samples are subjected to extensive
protease digestion with Bacillus sp
protease. This process releases amino
acids including glycated valines from the
hemoglobin beta chains. Glycated valines
then serve as substrates for specific
recombinant fructosyl valine oxidase
(FVO) enzyme, produced in E. coli. The
recombinant FVO specifically cleaves N-
terminal valines and produces hydrogen
peroxide. This, in turn, is measured using
a horse radish peroxidase (POD) catalyzed
reaction and a suitable chromagen. | Diazyme Direct HbA1c Assay is an
enzymatic assay in which lysed whole
blood samples are subjected to extensive
protease digestion with Bacillus sp
protease. This process releases amino
acids including glycated valines from the
hemoglobin beta chains. Glycated valines
then serve as substrates for specific
recombinant fructosyl valine oxidase
(FVO) enzyme. The recombinant FVO
specifically cleaves N-terminal valines
and produces hydrogen peroxide. This, in
turn, is measured using a horseradish
peroxidase (POD) catalyzed reaction and
a suitable chromogen. | Same |
| The HbA1c concentration is expressed
directly as %HbA1c by use of a suitable
calibration curve in which the calibrators
have values for each level in %HbA1c. | The HbA1c concentration is expressed
directly as %HbA1c by use of a suitable
calibration curve in which the
calibrators have values for each level in
%HbA1c. | |
Test Objective
| Direct HbA1c Enzymatic Assay
(K070734) | Direct HbA1c Assay (Enzymatic, On-
Board Lysis) | Equivalency |
|------------------------------------------------------------------------------------|------------------------------------------------------------------------------------|-------------|
| For the in vitro quantitative
determination of % HbA1c in human
whole blood. | For the in vitro quantitative
determination of % HbA1c in human
whole blood. | Same |
Type of Test
14
| Direct HbA1c Enzymatic Assay
(K070734) | Direct HbA1c Assay (Enzymatic, On-
Board Lysis) | Equivalency |
|-------------------------------------------|----------------------------------------------------|-------------|
| Quantitative | Quantitative | Same |
Specimen Type
| Direct HbA1c Enzymatic Assay
(K070734) | Direct HbA1c Assay (Enzymatic, On-
Board Lysis) | Equivalency |
|-------------------------------------------|----------------------------------------------------|-------------|
| EDTA Human whole blood. | K2 EDTA Human whole blood. | Same |
Specimen Lysis
| Direct HbA1c Enzymatic Assay
(K070734) | Direct HbA1c Assay (Enzymatic, On-
Board Lysis) | Equivalency |
|-------------------------------------------------------------|---------------------------------------------------------------|-------------|
| Human whole blood Specimens are
lysed manually off-board | Human whole blood specimens are
lysed on-board on analyzer | Different |
Product Type
| Direct HbA1c Enzymatic Assay
(K070734) | Direct HbA1c Assay (Enzymatic, On-
Board Lysis) | Equivalency |
|--------------------------------------------------------|--------------------------------------------------------|-------------|
| 3 Reagents plus lysis buffer, Calibrators,
Controls | 2 Reagents plus lysis buffer, Calibrators,
Controls | Different |
Performance Comparison
| Direct HbA1c Enzymatic Assay
(K070734) | Direct HbA1c Assay (Enzymatic, On-
Board Lysis) | Equivalency |
|---------------------------------------------------------------------------|--------------------------------------------------------------------------------|--------------------------|
| Linear Range: 4-16% HbA1c1
Total Precision: 1.8%
Method Comparison: | Linear Range: 4-12 % HbA1c1
Total Precision: 0.6-1.9%
Method Comparison: | Substantially
similar |
| Correlation Coefficient: 0.9874
Slope/Intercept: y = 1.0212x + 0.0135 | Correlation Coefficient: 0.9937
Slope/Intercept: y = 1.023x - 0.09 | |
¹Linear Range limited to 4-12% HbA1c so that the upper limit of AMR is not too far from NGSP protocol for certification sample range.
15
Summary of Rationale for Considering the Device Substantially Equivalent to Devices Approved for Interstate Commerce
In summary, the Intended Use of the modified device has not changed. The assay principle and fundamental scientific technology of the modified device has not changed. The major changes are in the number of reagents supplied (from 3 reagent plus lysis buffer to 2 reagents plus lysis buffer), Concentration of components in the reagents and labeling changes reflecting the modification. Design control activities including precision, limit of quantitation, linearity, accuracy and interference studies demonstrates that the Diazyme Direct HbA1c Assay (Enzymatic, On-Board Lysis) is substantially similar to the predicate device (K070734) and is safe and effective. There is no significant deviation between performance of the modified device Diazyme Direct HbA1c Assay (Enzymatic, On-Board Lysis) and the predicate device. The differences in the assay steps should not affect the safety and effectiveness of the Diazyme Direct Hbalc assay (Enzymatic, On-Board Lysis). Diazyme Direct HbA1c assay calibrator is traceable to NGSP certification for the modified device. The legally marketed device Tosoh HPLC HbA1c Test is used as standard NGSP certification method in NGSP reference laboratories and was thus selected for accuracy study. The Tosoh HPLC HbA1c Test is also the device used in accuracy studies for the predicate Diazyme HbA1c Enzymatic assay device (K070734). The accuracy studies with whole blood samples tested with the modified device unequivocally demonstrated excellent correlation. In conclusion, the Diazyme Direct HbA1c Assay (Enzymatic, On-Board Lysis) for measurement of HbA1c in patient whole blood samples is substantially equivalent to legally marketed devices. The differences in the assay steps should not affect the safety and effectiveness of the Diazyme Direct HbA1c assay (Enzymatic, On-Board Lysis) and risk analysis performed indicates no change in risk stance. The Diazyme Direct HbA1c assay gives similar diagnostic information as the other legally marketed HbA1c tests and offers users a rapid and convenient device for measuring HbA1c in human blood.