(119 days)
The D-10™ Hemoglobin A1c Program is intended for the quantitative determination of hemoglobin A1c (IFCC mmol/ mol and NGSP %) in human whole blood using ion- exchange high-performance liquid chromatography (HPLC) on the D-10TM Hemoglobin Testing System.
Hemoglobin Alc measurements are used as an aid in diagnosis of diabetes, as an aid to identify patients who may be at risk for developing diabetes mellitus, and for the monitoring of long-term blood glucose control in individuals with diabetes mellitus.
The D-10™ Hemoglobin A1c Program is intended for professional in vitro diagnostic use only.
The D-10™ Hemoglobin Testing System utilizes the principles of ion-exchange highperformance liquid chromatography (HPLC). A dual-piston, low pulsation HPLC pump and a proportioning value deliver the buffer solution to an analytical cartridge and detector. Whole blood samples undergo an automatic two step dilution process and then introduced into the analytical flow path. Pre-diluted samples are aspirated directly and introduced into the analytical flow path. Between sample injections, the sample probe is rinsed with Wash/Diluent Solution to minimize sample carryover.
A programmed buffer gradient of increasing ionic strength delivers the sample to the analytical cartridge where the hemoglobin species are separated based upon their ionic interactions with the cartridge material and the buffer gradient. The separated hemoglobin species then pass through the photometer flow cell where changes in the absorbance are measured at 415 nm and recorded as a digital chromatogram.
The software performs a reduction of raw data collected from each analysis that may indicate use of a calibration factor. A samples report and chromatogram are generated for each sample.
The D-10™ Hemoglobin A1c Program is designed to be used on the D-10™ Hemoglobin Testing System with or without a D-10 Rack Loader.
The provided text describes a 510(k) premarket notification for the D-10 Hemoglobin A1c Program. This device is an in vitro diagnostic (IVD) used for the quantitative determination of hemoglobin A1c (HbA1c) in human whole blood using ion-exchange high-performance liquid chromatography (HPLC).
Here's an analysis of the acceptance criteria and the study that proves the device meets those criteria, based on the provided document:
1. Table of Acceptance Criteria and Reported Device Performance:
The document doesn't explicitly state "acceptance criteria" in a single table, but rather presents performance characteristics derived from various studies. Based on the "Summary of Nonclinical Performance Data" and the conclusion that "The performance criteria as stipulated by the Special Controls requirements for HbA1c systems that diagnose diabetes have clearly been met," we can infer the acceptance criteria from the reported results. The critical performance metrics for an HbA1c assay for diabetes diagnosis typically include precision (CV%), linearity, and method comparison (bias).
| Performance Characteristic | Acceptance Criteria (Inferred from successful study results and regulatory requirements) | Reported Device Performance (D-10™ Hemoglobin A1c Program) |
|---|---|---|
| Precision (NGSP %) | Typically, CV% values are expected to be low, especially at clinical decision points. (Implicitly, the results demonstrate acceptable precision as per CLSI EP05-A2 guidelines). | Total Precision (Combined Instruments): Patient 1 (5.2%): 2.0% CV Patient 2 (6.7%): 1.6% CV Patient 3 (8.3%): 1.6% CV Patient 4 (12.7%): 2.2% CV Control 1 (5.6%): 1.7% CV Control 2 (10.3%): 2.2% CV QC 1 (5.5%): 2.0% CV QC 2 (9.9%): 1.8% CV QC 3 (15.4%): 2.2% CV |
| Precision (IFCC mmol/mol) | Similarly, low CV% values are expected. | Total Precision (Combined Instruments): Patient 1 (33 mmol/mol): 3.4% CV Patient 2 (50 mmol/mol): 2.3% CV Patient 3 (67 mmol/mol): 2.1% CV Patient 4 (115 mmol/mol): 2.7% CV Control 1 (37 mmol/mol): 2.8% CV Control 2 (89 mmol/mol): 2.7% CV QC 1 (37 mmol/mol): 3.2% CV QC 2 (85 mmol/mol): 2.3% CV QC 3 (145 mmol/mol): 2.5% CV |
| Linearity (NGSP %) | Maximum measured difference of ±0.1% between predicted 1st and 2nd order results across the reportable range. | Linear from 3.9 – 18.8% HbA1c, with a maximum measured difference of ±0.1%. |
| Linearity (IFCC mmol/mol) | Maximum measured difference of ±1 mmol/mol across the reportable range. | Linear from 19 – 182 mmol/mol, with a maximum measured difference of ±1 mmol/mol. |
| Method Comparison (Bias vs. NGSP SRL) | Biases at clinical decision levels should be within acceptable limits for diagnostic accuracy. (Implicitly, the reported biases are considered acceptable). | Bias Estimation: 5.0±0.5% HbA1c: -0.05% (-0.96% Bias) 6.5±0.5% HbA1c: 0.00% (0.03% Bias) 8.0±0.5% HbA1c: -0.08% (-0.98% Bias) 12.0±1.0% HbA1c: -0.10% (-0.87% Bias) |
| Total Error (TE) at Decision Levels | Should be below a specified threshold (e.g., as defined by CLIA or other regulatory guidelines for HbA1c testing). | Total Error Estimation: 5.0% A1c: 4.9% TE 6.5% A1c: 3.2% TE 8.0% A1c: 4.1% TE 12.0% A1c: 5.2% TE |
| Endogenous Interference | No significant interference (defined as a ± 7% change in %HbA1c value from the control) up to stated concentrations. | No significant interference observed for Lipemia (6000 mg/dL), Conjugated bilirubin (60 mg/dL), Unconjugated bilirubin (60 mg/dL), Glucose (2000 mg/dL), Rheumatoid factor (750 IU/mL), Total protein (21 g/dL). |
| Drug Interference | No significant interference (defined as a more than ± 7% change in %HbA1c value from the control) at therapeutic levels up to stated concentrations. | No significant interference observed for 15 common drugs (e.g., Acetylcysteine, Ampicillin-Na, Ascorbic acid, Cefoxitin, etc.) at specified concentrations. |
| Cross Reactivity with Hemoglobin Derivatives | No significant interference (defined as more than a ±7% change in HbA1c value from the control) at physiological levels. | No significant interference for Acetylated Hb (up to 49 mg/dL), Carbamylated Hb (up to 3.5%), and Labile A1c (up to 6%). |
| Hemoglobin Variant Interference | Acceptable bias for common variants (HbS, C, D, E, A2) and no significant interference for HbF up to 10%. | No significant interference for HbC (≤ 40%), HbD (≤ 43%), HbS (≤ 43%), HbE (≤ 32%), HbA2 (≤ 6%), and HbF (≤ 10%). Device has significant positive interference with HbF > 10%, rendering results invalid. |
| Traceability/Standardization | Traceable to IFCC reference calibrators and NGSP certified. | Traceable to IFCC reference calibrators. NGSP certified (certification expires annually). |
2. Sample Sizes Used for the Test Set and Data Provenance:
- Precision/Reproducibility: Four EDTA whole blood samples at target HbA1c concentrations (~5%, ~6.5%, ~8%, ~12%) and five quality control materials were used. Each sample/control was run in duplicate, in 2 runs per day, on 3 instruments for 20 days, and repeated with 3 different kit lots, yielding a total of 720 results per sample over 60 days. The data provenance is not explicitly stated (e.g., country of origin, retrospective/prospective clinical samples), but it implies prospective collection for the study.
- Linearity: Low (3.9% HbA1c) and high (18.8% HbA1c) EDTA whole blood patient samples were mixed in varying ratios to create 11 sample pools. The exact number of individual patient samples generating the pools is not specified.
- Method Comparison: 128 variant-free whole blood K3 EDTA samples were evaluated. The range of HbA1c was from 3.9% to approximately 19.0% (19 to 184 mmol/mol). Data provenance is not explicitly stated for these samples, but they are referred to as "patient samples."
- Endogenous/Drug Interference & Hemoglobin Derivatives Interference: Two EDTA whole blood sample pools (low level ~6.5% HbA1c and high level ~8.0% HbA1c) were used for spiking experiments. Ten replicates of each pool with interferent and control were analyzed. The specific number of individual patient samples that formed these pools is not detailed.
- Hemoglobin Variant Study: A panel of normal and diabetic whole blood EDTA patent variant samples for HbS (n=22), HbC (n=20), HbD (n=22), HbE (n=23), HbA2 (n=20), and HbF (n=24) were used. These are patient samples with known variants.
3. Number of Experts Used to Establish the Ground Truth for the Test Set and Qualifications of Those Experts:
- For Method Comparison: The D-10™ Hemoglobin A1c Program results were compared to "testing performed at a secondary NGSP SRL reference laboratory using a cleared HPLC-based HbA1c assay." The NGSP SRL (National Glycohemoglobin Standardization Program Secondary Reference Lab) is itself a highly qualified reference standard, implying expert oversight and standardized methods for ground truth determination, though specific human experts are not named or qualified in this document.
- For Hemoglobin Variant Study: Comparison was made to a "NGSP reference method that has been demonstrated to be free from the hemoglobin interferent." Similar to the method comparison, this relies on a standardized reference method rather than individual expert adjudication for each sample.
4. Adjudication Method for the Test Set:
Not applicable in the traditional sense of image or clinical outcome adjudication by multiple experts. The ground truth for the method comparison and variant studies was established by comparison to a standardized NGSP reference method, which inherently provides a highly controlled and validated "truth" for HbA1c measurements.
5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study was Done:
No, a MRMC comparative effectiveness study was not done. This device is an in vitro diagnostic (IVD) for laboratory use, not typically subject to human reader interpretation or MRMC studies.
6. If a Standalone (i.e., algorithm only without human-in-the-loop performance) Was Done:
Yes, the studies described are all standalone performance evaluations of the D-10™ Hemoglobin A1c Program device. The tests, such as precision, linearity, method comparison, and interference studies, assess the analytical performance of the algorithm and hardware without human interpretation in the loop determining the HbA1c value.
7. The Type of Ground Truth Used:
The ground truth for the performance studies primarily relies on:
- Reference Methods: For method comparison and variant studies, the ground truth was established by a "secondary NGSP SRL reference laboratory using a cleared HPLC-based HbA1c assay" and an "NGSP reference method."
- Expected Values/Spiking: For precision, linearity, and interference studies, ground truth was based on preparing samples with known concentrations or by comparing to control samples without interferents.
- Consensus/Certification: The general standardization is traced to IFCC reference calibrators and NGSP certification.
8. The Sample Size for the Training Set:
This document describes premarket notification studies for an in vitro diagnostic device, not a machine learning algorithm. Therefore, there is no explicit "training set" in the context of machine learning model development. The device relies on established chemical and physical principles (ion-exchange HPLC) with programmed parameters, rather than learning from a large dataset. Calibration of the instrument is performed using calibrator materials, but this is distinct from a machine learning training set.
9. How the Ground Truth for the Training Set Was Established:
As there is no machine learning-based training set, this question is not applicable. The device's operational parameters and calibration are based on established analytical chemistry principles and reference materials. The value assignment for the calibrator materials "were previously reviewed under 510(k) submission K031043," indicating their ground truth was established through a separate, regulated process.
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Image /page/0/Picture/1 description: The image is a black and white logo for the Department of Health & Human Services - USA. The logo is circular, with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" arranged around the perimeter of the circle. In the center of the circle is a stylized image of three human profiles facing to the right.
Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002
BIO-RAD LABORATORIES, INC. JACKIE BUCKLEY REGULATORY AFFAIRS SUPERVISOR 4000 ALFRED NOBEL DR. HERCULES CA 94547
October 14, 2016
Re: K161687
Trade/Device Name: D-10 Hemoglobin A1c Program Regulation Number: 21 CFR 862.1373 Regulation Name: Glycosylated hemoglobin assay Regulatory Class: II Product Code: PDJ Dated: August 30, 2016 Received: September 1, 2016
Dear Ms. Buckley:
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 electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.
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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
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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
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Indications for Use
510(k) Number (if known) K161687
Device Name D-10TM Hemoglobin A1c Program
Indications for Use (Describe)
The D-10™ Hemoglobin A1c Program is intended for the quantitative determination of hemoglobin A1c (IFCC mmol/ mol and NGSP %) in human whole blood using ion- exchange high-performance liquid chromatography (HPLC) on the D-10TM Hemoglobin Testing System.
Hemoglobin Alc measurements are used as an aid in diagnosis of diabetes, as an aid to identify patients who may be at risk for developing diabetes mellitus, and for the monitoring of long-term blood glucose control in individuals with diabetes mellitus.
The D-10™ Hemoglobin A1c Program is intended for professional in vitro diagnostic use only.
Type of Use (Select one or both, as applicable)
| Prescription Use (Part 21 CFR 801 Subpart D) | |
| Over-The-Counter Use (21 CFR 801 Subpart C) |
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510(k) Summary (Summary of Safety and Effectiveness)
This Summary of 510(k) Safety and Effectiveness is being submitted in accordance with the requirements of SMDA 1990 and 21 CFR 807.92.
The assigned 510(k) number is: K161687
Date Summary prepared: Oct. 12, 2016
1. Applicant Name:
Bio-Rad Laboratories, Inc. Clinical Diagnostics Group 4000 Alfred Nobel Drive Hercules, California 94547
2. Contact Person(s):
Jackie Buckley, Regulatory Affairs Supervisor Telephone Number: (510) 741-5309 FAX: (510) 741-6471 E-Mail: jackie buckley@bio-rad.com
Alfred Evans, RA/QA Director Telephone Number: (510) 741-4579 FAX: (510) 741-6471 E-Mail:al evans@bio-rad.com
3. Device Name/Trade Name:
Reagents: Trade Name: D-10™ Hemoglobin A1c Program Classification Name: Hemoglobin A1c Test System Common Name: HbA1c Product Code: PDJ C.F.R Section: 21 CFR 862.1373 Device classification: Class II Panel Classification: Chemistry
4. Predicate Device:
| Predicate DeviceName | Predicate Device510(k) Number |
|---|---|
| VARIANT II TURBO HbA1c Kit -2.0 | K142448 |
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5. Description of the Device:
The D-10™ Hemoglobin Testing System utilizes the principles of ion-exchange highperformance liquid chromatography (HPLC). A dual-piston, low pulsation HPLC pump and a proportioning value deliver the buffer solution to an analytical cartridge and detector. Whole blood samples undergo an automatic two step dilution process and then introduced into the analytical flow path. Pre-diluted samples are aspirated directly and introduced into the analytical flow path. Between sample injections, the sample probe is rinsed with Wash/Diluent Solution to minimize sample carryover.
A programmed buffer gradient of increasing ionic strength delivers the sample to the analytical cartridge where the hemoglobin species are separated based upon their ionic interactions with the cartridge material and the buffer gradient. The separated hemoglobin species then pass through the photometer flow cell where changes in the absorbance are measured at 415 nm and recorded as a digital chromatogram.
The software performs a reduction of raw data collected from each analysis that may indicate use of a calibration factor. A samples report and chromatogram are generated for each sample.
The D-10™ Hemoglobin A1c Program is designed to be used on the D-10™ Hemoglobin Testing System with or without a D-10 Rack Loader.
Reagents:
The D-10™ HbA1c reagents contain the following components: Description
D-10 ™ Hemoglobin Are Analytical Cartridge. Cation exchange cartridge (400 tests), 4.0 mm ID x 30 mm.
D-10 ™ Wash/Diluent Solution. Each bottle contains 1600 mL of deionized water with <0.05 sodium azide as a preservative.
Floppy diskette with D-10 Hemoglobin A1c "Update Kit" program parameters.
D-10™ Hemoglobin Are Elution Buffer 1. Each bottle contains 2000 mL of a Bis-Tris/Phosphate buffer. Contains <0.05% sodium azide as a preservative.
D-10 "" Hemoglobin Are Elution Buffer 2. Each bottle contains 1000 mL of a Bis-Tris/Phosphate buffer. Contains<0.05% sodium azide as a preservative.
D-10 10 10 Hemoglobin A16 Calibrator/Diluent Set. One set consisting of 3 vials of Calibrator 1, 3 vials of Calibrator 2, and 1 bottle of Calibrator Diluent. The calibrator vials contain Ivophilized human red blood cell hemolysate with gentamicin, tobramycin, and EDTA as preservative. Reconstituted volume is 7mL per vial.
Calibrator Diluent contains 100 mL of deionized water with <0.05% sodium azide as preservative.
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CD-ROM with D-10 Hemoglobin A1c "Update Kit" program parameters
Whole Blood Primer. Each vial contains lyophilized human red blood cell hemolysate with gentamicin, tobramycin, and EDTA as preservatives. Reconstituted volume is 1.0 mL per vial.
Sample Vials. 100 polypropylene microvials with pierceable caps, 1.5mL
D-10 Hemoglobin A1c Supplemental Reagent Pack. This reagent pack is used as a supplement to the D-10 Hemoglobin A1c Reorder Pack.
6. Indications for Use:
The D-10™ Hemoglobin A1c Program is intended for the quantitative determination of hemoglobin A1c (IFCC mmol/mol and NGSP %) in human whole blood using ionexchange high-performance liquid chromatography (HPLC) on the D-10™ Hemoglobin Testing System.
Hemoqlobin A1c measurements are used as an aid in diagnosis of diabetes, as an aid to identify patients who may be at risk for developing diabetes mellitus, and for the monitoring of long-term blood glucose control in individuals with diabetes mellitus.
The D-10™ Hemoglobin A1c Program is intended for professional in vitro diagnostic use only.
7. Substantial Equivalence Information:
Predicate Device Information:
| Predicate Device | Predicate Device |
|---|---|
| Name | 510(k) Number |
| VARIANT II TURBO HbA1c Kit -2.0 | K142448 |
The comparison of the technological characterizes of the D-10 Hemoglobin A1c Program (candidate assay) utilizes principles of ion-exchange high-performance liquid chromatography (HPLC) similar to the same technology of the VARIANT II TURBO HbA1c Kit - 2.0 (predicate device).
Tables 1 and 2 provide the similarities and differences between the candidate assay and the predicate assay.
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| ReagentSimilarities and Differences | ||
|---|---|---|
| Features | Candidate Device:D-10™ Hemoglobin A1c(K)161687 | Predicate Device:VARIANT™ II TURBO HbA1cKit - 2.0(K)142448 |
| Intended Use | Same | Intended for thequantitative determination ofhemoglobin A1c (IFCC mmol/moland NGSP %) |
| Platform | D-10™ Hemoglobin TestingSystem | VARIANT™II TURBOHemoglobin Testing System andVARIANT™II TURBO LinkHemoglobin Testing System |
| MeasuringInterval | 3.9 to 18.8% (NSGP)19 – 182 mmol/molHbA1c (IFCC) | 3.4 to 20.6 % (NSGP)14 – 203 mmol/molHbA1c (IFCC) |
| Specimen Type | Same | Human Whole blood |
| Assay Principle | Same | Ion exchange HPLC |
| Matrices | K₂-EDTA, K₃-EDTA | K₂-EDTA, K₃-EDTAHemoglobin Capillary Collection Kit |
| Standardization | Same | Traceable to the DiabetesControl and Complications Trial(DCCT) reference method andIFCC. Certified via the NationalGlycohemoglobin StandardizationProgram (NGSP) |
Table 1: Reagent Similarities and Differences
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8. Summary of Nonclinical Performance Data:
Precision/Reproducibility: a.
The precision of the D-10™ Hemoglobin A1c Program was evaluated based on CLSI EP05-A2 guidelines, Evaluation of Precision Performance of Quantitative Measurement Methods using a modified study design. Four EDTA whole blood samples at the following targeted HbA1c concentrations of ~5%, ~6.5%, ~8% and ~12% were utilized in the study. In addition, five quality control materials were also tested. The samples were run in duplicate in 2 runs per day on 3 instruments for 20 days. The study was repeated using 3 different kit lots, yielding a total of 720 results per sample over a 60-day period. NGSP results are shown in Tables 2-5. IFCC results are shown in Tables 6-9.
| HbA1c,% | Between-Run | Between-Day | Between-Lot | Total Precision | |||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| Sample ID | Repeatability | SD | %CV | SD | %CV | SD | %CV | SD | %CV | ||
| Patient 1 | 5.2 | 0.07 | 1.3 | 0.00 | 0.0 | 0.04 | 0.7 | 0.08 | 1.6 | 0.11 | 2.2 |
| Patient 2 | 6.7 | 0.07 | 1.0 | 0.00 | 0.0 | 0.07 | 1.1 | 0.05 | 0.7 | 0.11 | 1.7 |
| Patient 3 | 8.4 | 0.05 | 0.6 | 0.01 | 0.2 | 0.08 | 1.0 | 0.02 | 0.2 | 0.10 | 1.2 |
| Patient 4 | 12.7 | 0.09 | 0.7 | 0.06 | 0.5 | 0.17 | 1.3 | 0.21 | 1.7 | 0.29 | 2.3 |
| Control 1 | 5.6 | 0.07 | 1.2 | 0.00 | 0.0 | 0.05 | 0.9 | 0.05 | 0.9 | 0.10 | 1.8 |
| Control 2 | 10.4 | 0.10 | 0.9 | 0.00 | 0.0 | 0.12 | 1.1 | 0.12 | 1.2 | 0.20 | 1.9 |
| QC 1 | 5.5 | 0.06 | 1.0 | 0.00 | 0.0 | 0.05 | 0.9 | 0.08 | 1.5 | 0.11 | 2.1 |
| QC 2 | 10.0 | 0.13 | 1.3 | 0.00 | 0.0 | 0.09 | 0.9 | 0.06 | 0.6 | 0.17 | 1.7 |
| QC 3 | 15.6 | 0.11 | 0.7 | 0.15 | 1.0 | 0.08 | 0.5 | 0.17 | 1.1 | 0.27 | 1.7 |
Table 2: Instrument 1 % CV by Sample (NGSP)
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| Sample ID | HbA1c,% | Repeatability | Between-Run | Between-Day | Between-Lot | TotalPrecision | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV | ||
| Patient 1 | 5.2 | 0.04 | 0.9 | 0.02 | 0.4 | 0.04 | 0.8 | 0.08 | 1.5 | 0.10 | 1.9 |
| Patient 2 | 6.6 | 0.03 | 0.5 | 0.04 | 0.6 | 0.05 | 0.8 | 0.02 | 0.3 | 0.08 | 1.2 |
| Patient 3 | 8.2 | 0.04 | 0.5 | 0.04 | 0.4 | 0.07 | 0.8 | 0.02 | 0.2 | 0.09 | 1.0 |
| Patient 4 | 12.5 | 0.06 | 0.5 | 0.08 | 0.6 | 0.13 | 1.1 | 0.21 | 1.7 | 0.27 | 2.1 |
| Control 1 | 5.6 | 0.04 | 0.8 | 0.02 | 0.3 | 0.06 | 1.0 | 0.07 | 1.2 | 0.10 | 1.7 |
| Control 2 | 10.1 | 0.04 | 0.4 | 0.06 | 0.6 | 0.08 | 0.8 | 0.09 | 0.9 | 0.14 | 1.4 |
| QC 1 | 5.5 | 0.03 | 0.6 | 0.02 | 0.4 | 0.06 | 1.0 | 0.08 | 1.5 | 0.11 | 1.9 |
| QC 2 | 9.8 | 0.07 | 0.7 | 0.06 | 0.6 | 0.09 | 0.9 | 0.04 | 0.4 | 0.13 | 1.4 |
| QC 3 | 15.2 | 0.07 | 0.4 | 0.08 | 0.5 | 0.14 | 0.9 | 0.24 | 1.6 | 0.29 | 1.9 |
Table 3: Instrument 2 % (CV by Sample (NGSP))
Table 4: Instrument 3 (% CV by Sample (NGSP))
| Between- | Between- | Total | |||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| Sample ID | HbA1c, | Repeatability | Run | Day | Between-Lot | Precision | |||||
| % | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV | |
| Patient 1 | 5.2 | 0.06 | 1.2 | 0.00 | 0.0 | 0.06 | 1.2 | 0.05 | 0.9 | 0.10 | 1.9 |
| Patient 2 | 6.7 | 0.05 | 0.7 | 0.02 | 0.3 | 0.08 | 1.2 | 0.00 | 0.0 | 0.09 | 1.4 |
| Patient 3 | 8.3 | 0.05 | 0.6 | 0.03 | 0.4 | 0.11 | 1.3 | 0.04 | 0.5 | 0.13 | 1.6 |
| Patient 4 | 12.7 | 0.07 | 0.6 | 0.07 | 0.6 | 0.17 | 1.3 | 0.21 | 1.6 | 0.29 | 2.2 |
| Control 1 | 5.6 | 0.05 | 0.9 | 0.00 | 0.0 | 0.07 | 1.2 | 0.04 | 0.8 | 0.10 | 1.7 |
| Control 2 | 10.3 | 0.08 | 0.8 | 0.05 | 0.5 | 0.14 | 1.3 | 0.08 | 0.8 | 0.19 | 1.8 |
| QC 1 | 5.5 | 0.04 | 0.8 | 0.02 | 0.3 | 0.07 | 1.2 | 0.07 | 1.3 | 0.11 | 2.0 |
| QC 2 | 9.9 | 0.09 | 0.9 | 0.00 | 0.0 | 0.12 | 1.2 | 0.13 | 1.3 | 0.20 | 2.0 |
| QC 3 | 15.5 | 0.10 | 0.6 | 0.11 | 0.7 | 0.18 | 1.1 | 0.07 | 0.4 | 0.24 | 1.6 |
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| Between- | Between- | Total | |||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Sample ID | HbA1c | Repeatability | Between-Run | Day | Instruments | Between-Lot | Precision | ||||||
| % | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV | |
| Patient 1 | 5.2 | 0.06 | 1.2 | 0.00 | 0.0 | 0.05 | 0.9 | 0.00 | 0.0 | 0.07 | 1.4 | 0.10 | 2.0 |
| Patient 2 | 6.7 | 0.05 | 0.8 | 0.01 | 0.2 | 0.07 | 1.0 | 0.05 | 0.8 | 0.03 | 0.5 | 0.11 | 1.6 |
| Patient 3 | 8.3 | 0.05 | 0.6 | 0.03 | 0.3 | 0.09 | 1.1 | 0.08 | 0.9 | 0.03 | 0.3 | 0.13 | 1.6 |
| Patient 4 | 12.7 | 0.08 | 0.6 | 0.07 | 0.6 | 0.16 | 1.2 | 0.00 | 0.0 | 0.21 | 1.7 | 0.28 | 2.2 |
| Control 1 | 5.6 | 0.06 | 1.0 | 0.00 | 0.0 | 0.06 | 1.0 | 0.00 | 0.0 | 0.05 | 0.9 | 0.10 | 1.7 |
| Control 2 | 10.3 | 0.08 | 0.7 | 0.04 | 0.4 | 0.12 | 1.1 | 0.14 | 1.3 | 0.10 | 1.0 | 0.22 | 2.2 |
| QC 1 | 5.5 | 0.05 | 0.8 | 0.00 | 0.1 | 0.06 | 1.0 | 0.00 | 0.0 | 0.08 | 1.4 | 0.11 | 2.0 |
| QC 2 | 9.9 | 0.10 | 1.0 | 0.00 | 0.0 | 0.10 | 1.0 | 0.05 | 0.5 | 0.09 | 0.9 | 0.17 | 1.8 |
| QC 3 | 15.4 | 0.09 | 0.6 | 0.12 | 0.8 | 0.14 | 0.9 | 0.20 | 1.3 | 0.17 | 1.1 | 0.34 | 2.2 |
Table 5: Instruments Combined (% CV by Sample (NGSP))
Table 6: Instrument 1 (% CV by Sample (IFCC Units- mmol/mol))
| Sample ID | HbA1c,mmol/mol | Repeatability | Between-Run | Between-Day | Between-Lot | Total Precision | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV | ||
| Patient 1 | 33 | 0.76 | 2.3 | 0.00 | 0.0 | 0.40 | 1.2 | 0.89 | 2.7 | 1.24 | 3.7 |
| Patient 2 | 50 | 0.76 | 1.5 | 0.00 | 0.0 | 0.81 | 1.6 | 0.54 | 1.1 | 1.23 | 2.5 |
| Patient 3 | 68 | 0.53 | 0.8 | 0.16 | 0.2 | 0.92 | 1.3 | 0.17 | 0.3 | 1.08 | 1.6 |
| Patient 4 | 116 | 1.01 | 0.9 | 0.65 | 0.6 | 1.85 | 1.6 | 2.31 | 2.0 | 3.19 | 2.8 |
| Control 1 | 38 | 0.77 | 2.0 | 0.00 | 0.0 | 0.56 | 1.5 | 0.53 | 1.4 | 1.09 | 2.9 |
| Control 2 | 91 | 1.08 | 1.2 | 0.00 | 0.0 | 1.31 | 1.4 | 1.34 | 1.5 | 2.16 | 2.4 |
| QC 1 | 37 | 0.63 | 1.7 | 0.00 | 0.0 | 0.53 | 1.4 | 0.93 | 2.5 | 1.25 | 3.4 |
| QC 2 | 86 | 1.46 | 1.7 | 0.00 | 0.0 | 0.96 | 1.1 | 0.61 | 0.7 | 1.85 | 2.2 |
| QC 3 | 147 | 1.25 | 0.9 | 1.67 | 1.1 | 0.92 | 0.6 | 1.90 | 1.3 | 2.96 | 2.0 |
{10}------------------------------------------------
| Sample ID | HbA1c,mmol/mol | Repeatability | Between-Run | Between-Day | Between-Lot | Total Precision | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV | ||
| Patient 1 | 33 | 0.49 | 1.5 | 0.20 | 0.6 | 0.43 | 1.3 | 0.84 | 2.6 | 1.08 | 3.3 |
| Patient 2 | 49 | 0.36 | 0.7 | 0.42 | 0.9 | 0.58 | 1.2 | 0.24 | 0.5 | 0.83 | 1.7 |
| Patient 3 | 66 | 0.40 | 0.6 | 0.38 | 0.6 | 0.72 | 1.1 | 0.19 | 0.3 | 0.93 | 1.4 |
| Patient 4 | 113 | 0.63 | 0.6 | 0.88 | 0.8 | 1.47 | 1.3 | 2.27 | 2.0 | 2.91 | 2.6 |
| Control 1 | 37 | 0.46 | 1.2 | 0.19 | 0.5 | 0.61 | 1.6 | 0.70 | 1.9 | 1.05 | 2.8 |
| Control 2 | 87 | 0.45 | 0.5 | 0.66 | 0.8 | 0.89 | 1.0 | 0.96 | 1.1 | 1.54 | 1.8 |
| QC 1 | 37 | 0.38 | 1.0 | 0.21 | 0.6 | 0.61 | 1.7 | 0.91 | 2.5 | 1.18 | 3.2 |
| QC 2 | 84 | 0.72 | 0.9 | 0.67 | 0.8 | 1.00 | 1.2 | 0.43 | 0.5 | 1.46 | 1.7 |
| QC 3 | 142 | 0.73 | 0.5 | 0.83 | 0.6 | 1.52 | 1.1 | 2.58 | 1.8 | 3.19 | 2.2 |
Table 7: Instrument 2 (% CV by Sample (IFCC Units- mmol/mol))
Table 8: Instrument 3 (% CV by Sample (IFCC Units- mmol/mol))
| HbA1c, | Repeatability | Between-Run | Between-Day | Between-Lot | Total Precision | ||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| Sample ID | mmol/mol | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| Patient 1 | 33 | 0.70 | 2.1 | 0.00 | 0.0 | 0.66 | 2.0 | 0.51 | 1.6 | 1.09 | 3.3 |
| Patient 2 | 50 | 0.51 | 1.0 | 0.22 | 0.4 | 0.85 | 1.7 | 0.00 | 0.0 | 1.02 | 2.0 |
| Patient 3 | 68 | 0.56 | 0.8 | 0.33 | 0.5 | 1.20 | 1.8 | 0.46 | 0.7 | 1.44 | 2.1 |
| Patient 4 | 115 | 0.80 | 0.7 | 0.80 | 0.7 | 1.81 | 1.6 | 2.26 | 2.0 | 3.11 | 2.7 |
| Control 1 | 37 | 0.57 | 1.5 | 0.00 | 0.0 | 0.73 | 2.0 | 0.46 | 1.2 | 1.03 | 2.8 |
| Control 2 | 89 | 0.87 | 1.0 | 0.60 | 0.7 | 1.51 | 1.7 | 0.93 | 1.0 | 2.07 | 2.3 |
| QC 1 | 36 | 0.45 | 1.2 | 0.18 | 0.5 | 0.73 | 2.0 | 0.77 | 2.1 | 1.16 | 3.2 |
| QC 2 | 85 | 1.00 | 1.2 | 0.00 | 0.0 | 1.35 | 1.6 | 1.44 | 1.7 | 2.21 | 2.6 |
| QC 3 | 145 | 1.05 | 0.7 | 1.24 | 0.9 | 1.93 | 1.3 | 0.74 | 0.5 | 2.63 | 1.8 |
{11}------------------------------------------------
| SampleID | HbA1c,mmol/mol | Repeatability | Between-Run | Between-Day | Between-Instruments | Between-Lot | TotalPrecision | ||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV | ||
| Patient 1 | 33 | 0.66 | 2.0 | 0.00 | 0.0 | 0.51 | 1.5 | 0.00 | 0.0 | 0.77 | 2.3 | 1.13 | 3.4 |
| Patient 2 | 50 | 0.57 | 1.1 | 0.13 | 0.3 | 0.76 | 1.5 | 0.58 | 1.2 | 0.34 | 0.7 | 1.17 | 2.3 |
| Patient 3 | 67 | 0.50 | 0.7 | 0.31 | 0.5 | 0.96 | 1.4 | 0.84 | 1.3 | 0.30 | 0.4 | 1.44 | 2.1 |
| Patient 4 | 115 | 0.83 | 0.7 | 0.78 | 0.7 | 1.72 | 1.5 | 0.00 | 0.0 | 2.29 | 2.0 | 3.08 | 2.7 |
| Control 1 | 37 | 0.60 | 1.6 | 0.00 | 0.0 | 0.63 | 1.7 | 0.00 | 0.0 | 0.57 | 1.5 | 1.04 | 2.8 |
| Control 2 | 89 | 0.84 | 0.9 | 0.48 | 0.5 | 1.26 | 1.4 | 1.49 | 1.7 | 1.09 | 1.2 | 2.44 | 2.7 |
| QC 1 | 37 | 0.50 | 1.4 | 0.04 | 0.1 | 0.63 | 1.7 | 0.00 | 0.0 | 0.88 | 2.4 | 1.20 | 3.2 |
| QC 2 | 85 | 1.10 | 1.3 | 0.00 | 0.0 | 1.12 | 1.3 | 0.59 | 0.7 | 0.93 | 1.1 | 1.92 | 2.3 |
| QC 3 | 145 | 1.04 | 0.7 | 1.29 | 0.9 | 1.52 | 1.0 | 2.22 | 1.5 | 1.90 | 1.3 | 3.69 | 2.5 |
Table 9: Instruments Combined (% CV by Sample (IFCC Units- mmol/mol))
b. Linearity
A linearity study was performed per CLSI EP06-A: Evaluation of the Linearity of Quantitative Measuring Procedures; A Statistical Approach. Linearity across the reportable range was performed using low (3.9%HbA1c) and high (18.8%HbA1c) EDTA whole blood patient samples. These samples were mixed together in varying ratios. The measured values were compared to the expected values based upon the dilution factor. Polynomial regression analysis (for first, second, and third order polynomials) were performed to determine the statistical significance of non-linearity. The higher order coefficients were found not to be significant and linearity was demonstrated.
% HbA1c (NGSP) using the D-10™ Hemoglobin A1c Program has demonstrated linearity from 3.9 – 18.8% HbA1c with the maximum measured difference of
± 0.1% between the predicted 1ª and 20 order results as shown in Table 10 below. Mmol/mol HbA1c (IFCC) has been demonstrated as linear from 19 – 182 mmol/mol with the maximum measured difference of ±1 mmol/mol as shown in Table 11 below.
Table 10: Results of Linearity Study (NGSP %)
| Sample Pool | Predicted 1st Order | Predicted 2nd Order | Difference |
|---|---|---|---|
| Level 1 | 3.7 | 3.8 | 0.1 |
| Level 2 | 5.2 | 5.3 | 0.1 |
| Level 3 | 6.8 | 6.8 | 0.0 |
| Level 4 | 8.3 | 8.2 | 0.1 |
| Level 5 | 9.8 | 9.7 | 0.1 |
| Level 6 | 11.3 | 11.2 | 0.1 |
{12}------------------------------------------------
| Level 7 | 12.8 | 12.7 | 0.1 |
|---|---|---|---|
| Level 8 | 14.3 | 14.3 | 0.0 |
| Level 9 | 15.8 | 15.8 | 0.0 |
| Level 10 | 17.3 | 17.3 | 0.0 |
| Level 11 | 18.8 | 18.9 | 0.1 |
Table 11: Results of Linearity Study (IFCC mmol/mol)
| Sample Pool | Predicted 1st Order | Predicted 2nd Order | Difference |
|---|---|---|---|
| Level 1 | 17 | 18 | 1 |
| Level 2 | 34 | 34 | 0 |
| Level 3 | 50 | 50 | 0 |
| Level 4 | 67 | 67 | 0 |
| Level 5 | 83 | 83 | 0 |
| Level 6 | 100 | 99 | 1 |
| Level 7 | 116 | 116 | 0 |
| Level 8 | 133 | 133 | 0 |
| Level 9 | 149 | 149 | 0 |
| Level 10 | 166 | 166 | 0 |
| Level 11 | 182 | 183 | 1 |
Method Comparison ﻥ
A Method comparison study was performed per CLSI EP09-A3, Measurement Procedure Comparison and Bias Estimation Using Patient Samples. 128 variant-free whole blood K3 EDTA samples ranging from 3.9% to approximately 19.0 % (19 to approximately 184 mmol/mol) HbA1c were evaluated using the D-10™ Hemoglobin A1c Program on the D-10™ Hemoglobin Testing System. Samples were tested in a single determination over a 4 day period. The results were compared to testing performed at a secondary NGSP SRL reference laboratory using a cleared HPLC-based HbA1c assay. The distribution of samples spanned the measuring interval listed in Table 12.
| Hemoglobin A1c level | n | % Samples tested |
|---|---|---|
| ≤ 5% | 4 | 3.1 |
| 5 - 6% | 19 | 14.8 |
| 6 – 6.5% | 33 | 25.8 |
| 6.5 - 7% | 33 | 25.8 |
Table 12: Distribution of samples
{13}------------------------------------------------
| 7 – 8% | 20 | 15.6 |
|---|---|---|
| 8 – 9% | 10 | 7.8 |
| > 9% | 9 | 7.0 |
| Total samples | 128 | 100 |
Linear, Deming (weighted) and Passing-Bablok regression analyses were performed for the D-10™ Hemoglobin A1c versus the NGSP SRL reference method. Deming (weighted), Passing-Bablok and Linear regression analyses were performed for the D-10™ Hemoglobin A1c on the D-10 Hemoglobin Testing System versus the reference G8 HPLC method are summarized in Table 13.
| Table 13: Summary of Method Comparison Results | |||||
|---|---|---|---|---|---|
| Slope | 95% CI | y-Intercept | 95% CI | R² | |
| Linear | 0.9701 | 0.9587 – 0.9816 | 0.1801 | 0.0980 – 0.2622 | 0.9955 |
| Deming | 0.9722 | 0.9579 - 0.9866 | 0.1654 | 0.0637 - 0.2670 | 1.0000 |
| Passing-Bablok | 1.0000 | 1.0000 - 1.0000 | 0.0000 | 0.0000 - 0.0000 | 1.0000 |
Table 13: Summary of Method Comparison Results
{14}------------------------------------------------
Image /page/14/Figure/0 description: This image is a scatter plot titled "Scatter Plot with Deming Fit". The plot shows the relationship between NGSP on the x-axis and D-10 Hemoglobin A1c Program on the y-axis. The plot includes a Deming fit line represented by the equation (0.17 + 0.97x), along with 95% confidence interval bands and an identity line for reference. The data points are clustered tightly around the Deming fit line, indicating a strong correlation between the two variables.
Figure 1: Scatter Plot using Deming Fit, %HbA1c, NGSP SRL vs. D-10 Hemoglobin A1c.
- (1) The following biases between D-10 Hemoglobin A1c versus NGSP SRL Method (Reference method) were observed in Table 14.
| % HbA1c – DecisionLevel | Bias | % Bias |
|---|---|---|
| 5.0±0.5 | -0.05 | -0.96 |
| 6.5±0.5 | 0.00 | 0.03 |
| 8.0±0.5 | -0.08 | -0.98 |
| 12.0±1.0 | -0.10 | -0.87 |
Table 14: Bias Estimation
Total Error Decision Levels
Using the results of bias estimation (%Bias) in the method comparison study and precision estimates in the reproducibility study, Total Error (TE) at four concentrations: (5.0 %, 6.5%, 8.0% and 12.0%) were calculated as follows: %TE=|%Bias|+1.96 CV (1 + %Bias). The results are presented in Table 15.
{15}------------------------------------------------
| % A1c - Decision Level | % Bias | % CV | % TE |
|---|---|---|---|
| 5.0 | 0.96 | 2.0 | 4.9 |
| 6.5 | 0.03 | 1.6 | 3.2 |
| 8.0 | 0.98 | 1.6 | 4.1 |
| 12.0 | 0.87 | 2.2 | 5.2 |
Table 15: Total Error Estimation
d. Traceability, Stability, Expected Values (calibrators)
The D-10 Hemoglobin A1c test standardization is traceable to the International Federation of Clinical Chemistry (IFCC) reference calibrators. The D-10 Hemoglobin A1c assay is NGSP certified. The NGSP certification expires in one year. See NGSP website for current certification at http://www.ngsp.org. The derived results of (%) from the NGSP correlation are calculated from the individual quantitative results for Hemogloblin A1c (HbA1c). The International Federation of Clinical Chemistry (IFCC) units of mmol/mol are calculated using the Master Equation NGSP (%) = 0.09148 x IFCC (mmol/mol) + 2.152. HbA1c results are provided to the customers using two different units: NGSP equivalent units (%) and IFCC equivalent units (mmol/mol).
Calibrator Materials:
Value assignment for D-10™ Hemoglobin A1c Calibrators which are recommended for use with this device, were previously reviewed under 510(k) submission K031043.
e. Analytical specificity:
- Endogenous Interference i.)
An Endogenous Interference study was performed per CLSI EP07-A2, Interference Testing in Clinical Chemistry. Two EDTA whole blood sample pools were evaluated using a low level whole blood sample with a concentration ~6.5%HbA1c and a high level whole blood sample with a concentration of HbA1c of ~8.0%.
Coniugated bilirubin, unconiugated bilirubin and glucose, available in pure form, were obtained and stock solutions prepared at 10x the intended test concentration. The 10x stock solution of the test substance was pipetted into a low whole blood sample pool (at ~6.5% HbA1c) and a high whole blood sample pool (~8.0% HbA1c), making the test pool. Ten replicates of each pool prepared with the test and control samples were analyzed using the D-10TM Hemoglobin A1c on the D-10™Hemoglobin Testing System.
Rheumatoid factor, lipemia and total protein were not available as pure standards therefore serum samples with known concentration of these compounds were used. The test pool was prepared by mixing the serum sample known to have a high test substance concentration with a whole blood non-variant sample such that the concentration of test substance in the final mixture would be at the desired level. Ten replicates of each pool prepared with the test and control samples were analyzed using the D-10™ Hemoglobin A1c on the D-10™ Hemoglobin Testing System.
Significant interference was defined as a ± 7% chanqe in %HbA1c value from the control. Results in Table 16 showed no significant interference up to the stated concentrations.
{16}------------------------------------------------
| Concentration | ||
|---|---|---|
| Endogenous substance | Conventional (US)units | SI Units |
| Lipemia (Intralipid) | 6000 mg/dL | 60 g/L |
| Conjugated bilirubin | 60 mg/dL | 712 µmol/L |
| Unconjugated bilirubin | 60 mg/dL | 1026 µmol/L |
| Glucose | 2000 mg/dL | 111 mmol/L |
| Rheumatoid factor | 750 IU/mL | 750 kIU/mL |
| Total protein | 21 g/dL | 210 g/L |
Table 16: Endogenous Interference Study Results
ii.) Drug Interference:
A Drug Interference study was performed based per CLSI EP07-A2, Interference Testing in Clinical Chemistry. Two EDTA whole blood sample pools were evaluated using a low level whole blood sample with a concentration ~6.5%HbA1c and a high level whole blood sample with a concentration of ~8.0%HbA1c. Test samples were prepared by spiking each drug at the interferent concentration shown in Table 18. Ten replicates of each drug prepared with the test and control samples were analyzed using the D-10™ Hemoglobin A1c on the D-10™ Hemoglobin Testing System.
Significant interference was defined as a more than ± 7% change in %HbA1c value from the control. No significant interference was observed at therapeutic levels up to the stated concentrations in Table 17 on the following page.
{17}------------------------------------------------
| Potential DrugInterferent | Highest Level Tested showing no SignificantInterference | |
|---|---|---|
| Conventional (US)units | SI units | |
| Acetylcysteine | 166 mg/dL | 10.2 mmol/L |
| Ampicillin-Na | 1000 mg/dL | 28.65 mmol/L |
| Ascorbic acid | 300 mg/dL | 17.05 mmol/L |
| Cefoxitin | 2500 mg/dL | 58.55 mmol/L |
| Heparin | 5000 U/L | 5000 U/L |
| Levodopa | 20 mg/dL | 1015 µmol/L |
| Methyldopa | 20 mg/dL | 948 µmol/L |
| Metronidazole | 200 mg/dL | 11.7 mmol/L |
| Doxycyclin | 50 mg/dL | 1124 µmol/L |
| Acetylsalicylic acid | 1000 mg/dL | 55.51 mmol/L |
| Rifampicin | 64 mg/L | 78 µmol/L |
| Cyclosporine | 5 mg/L | 4 µmol/L |
| Acetaminophen | 200 mg/L | 1323 µmol/L |
| Ibuprofen | 500 mg/L | 2427 µmol/L |
| Theophylline | 100 mg/L | 556 µmol/L |
| Phenylbutazone | 400 mg/L | 1299 µmol/L |
Table 17: Drug Interference Study Results
iii.) Cross Reactivity with Hemoglobin Derivatives:
A Hemoglobin Derivatives Interference study was performed based on CLSI EP07-A2, Interference Testing in Clinical Chemistry. Potential interference from Acetylated hemogloblin (Hb), Carbamylated hemoglobin (Hb) and Labile HbA1c were evaluated using a low level whole blood EDTA sample with a concentration ~6.5%HbA1c and a high level whole blood EDTA sample with a concentration of
{18}------------------------------------------------
~8.0% HbA1c. The potentially interfering hemoglobin derivatives were spiked into the low and high level blood samples and each sample was analyzed using ten replicates each in the same analytical run on the D-10™ Hemoglobin Testing System with the D-10™Hemoglobin A1c.
Significant interference was defined as more than a ±7% change in HbA1c value from the control. The test result conclusions are as follows:
- . Acetylated Hb- up to 49 mg/dL does not interfere with this assay.
- . Carbamylated Hb - up to 3.5% (or 10 mg/dL potassium cyanate) does not interfere with this assay.
- Labile A1c- up to 6% (or 1000 mg/dL glucose) does not interfere with . this assay.
Results showed there was no cross reactivity with these substances at physiological levels.
- iv.) Hemoglobin Variant Study:
A Hemoglobin Variant study was performed using a panel of normal and diabetic whole blood EDTA patent variant samples known to contain hemoglobin variants S, C, E, D, A2 and F. Testing of the samples containing hemoglobin variants S, C, E, D, A2 and F were performed in duplicate using the D-10™ Hemoglobin A1c on the D-10™ Hemoglobin Testing System and compared to results obtained by a NGSP reference method that has been demonstrated to be free from the hemoglobin interferent. Table 18 contains the number of samples, range of samples and concentration of samples used in the Hemoglobin Variant Study. Table 19 contains the results for the Hemoglobin Variant study bias.
| Table 10: Variant samples used in Hemoglobin Variant Study | |||
|---|---|---|---|
| Hemoglobin | n | Range in % Abnormal | Range in %HbA1c |
| Variant | Variant | Concentration | |
| HbS | 22 | 31 - 43 | 5.6 - 11.5 |
| HbC | 20 | 31 - 40 | 5.0 - 10.7 |
| HbD | 22 | 35 - 43 | 5.8 - 10.0 |
| HbE | 23 | 21 - 32 | 5.9 - 11.6 |
| HbA2 | 20 | 5.0 - 6.2 | 5.0 - 14.5 |
| HbF | 24 | 3.3 - 32.6 | 4.7 - 14.4 |
Table 18: Variant samples used in Hemoglobin Variant Study
Table 19: Hemoqlobin Variant Study Bias Results
| HemoglobinVariant | Relative % Bias to Comparative Method | |
|---|---|---|
| Relative %Bias (Range of %Bias) forHbA1c | Relative %Bias (Range of %Bias)for HbA1c | |
| ~6.5% | ~9.0% | |
| HbS | 1.1 (-4.7 to 4.9) | 0.2 (-6.1 to5.3) |
| HbC | 2.6 (1.4 to 5.9) | -0.9 (-2.8 to 0.9) |
| HbD | -0.6 (-3.1 to 4.7) | 1.7 (-3.6 to 5.1) |
| HbE | 0.5 (-3.3 to 6.2) | 2.8 (1.2 to 5.2) |
| HbA2 | 0.6 (-1.8 to 1.8) | -1.7 (-2.8 to -0.7) |
| HbF | 3.1 (-1.5 to 8.8) | -0.6 (-2.5 to 4.8) |
{19}------------------------------------------------
This device has significant positive interference with fetal hemoglobin (HbF). HbA1c results are invalid for patients with abnormal amounts of HbF including those with known Hereditary Persistence of Fetal Hemoglobin.
Hemoglobin F concentrations up to 10% do not interfere with the test. Any sample with HbF>10% may result in higher than expected HbA1c values. Any sample with HbF >5% should be suspected of having a hemoglobinopathy.
No significant interference was observed for HbC (≤ 40%), HbD (≤ 43%), HbS (≤ 43%), HbE (≤ 32%),HbA2 (≤ 6%), and HbF(≤ 10%) at the concentrations tested in this study.
f. Matrix comparison
The data supports the use of the following blood collection tubes with the D-10™ Hemoglobin A1c test in Table 20.
| Table 20: Anticoagulant | |||
|---|---|---|---|
| 1 นิ้มเป็นครั้ง | |||
| Ko-EDTA | |||
| Kz.EDTA |
g. Expected Values/Reference Range
Hemoglobin A1c expected values range was cited from American Diabetes Association Standards of Medical Care in Diabetes 2014, 37 (Supplement 1) and American Diabetes Association. Standards of Medical Care in Diabetes - 2016 are presented in Table 21.
| Table 21: Hemoqlobin A1c Expected Values | ||
|---|---|---|
| Hemoglobin A1c | Suggested Diagnosis | |
|---|---|---|
| NGSP % | IFCC mmol/mol | |
| >6.5 | >47 | Diabetic |
| 5.7 - 6.4 | 39-46 | Pre-Diabetic |
| <5.7 | <39 | Non-Diabetic |
Conclusion:
The information and data in this 510(k) document demonstrate that the D-10™ Hemoglobin A1c Program as performed on the D-10™Hemoglobin Testing System is an accurate, reliable, precise test that correlates well with current cleared methods and NGSP standardized testing for the quantitation of HbA1c. The contents of this submission demonstrates that the D-10™ Hemoglobin A1c Program as performed on the D-10™ Hemoglobin Testing System is substantially equivalent to its predicate device, VARIANT II TURBO HbA1c Kit -2.0 and, therefore, safe and effective for its intended use. The performance criteria as stipulated by the Special Controls requirements for HbA1c systems that diagnose diabetes have clearly been met. The D-10™ Hemoglobin A1c Program must be found to be substantially equivalent to the predicate and, therefore, cleared by the agency for the intended use requested.
§ 862.1373 Hemoglobin A1c test system.
(a)
Identification. A hemoglobin A1c test system is a device used to measure the percentage concentration of hemoglobin A1c in blood. Measurement of hemoglobin A1c is used as an aid in the diagnosis of diabetes mellitus and as an aid in the identification of patients at risk for developing diabetes mellitus.(b)
Classification. Class II (special controls). The special controls for this device are:(1) The device must have initial and annual standardization verification by a certifying glycohemoglobin standardization organization deemed acceptable by FDA.
(2) The premarket notification submission must include performance testing to evaluate precision, accuracy, linearity, and interference, including the following:
(i) Performance testing of device precision must, at a minimum, use blood samples with concentrations near 5.0 percent, 6.5 percent, 8.0 percent, and 12 percent hemoglobin A1c. This testing must evaluate precision over a minimum of 20 days using at least three lots of the device and three instruments, as applicable.
(ii) Performance testing of device accuracy must include a minimum of 120 blood samples that span the measuring interval of the device and compare results of the new device to results of a standardized test method. Results must demonstrate little or no bias versus the standardized method.
(iii) Total error of the new device must be evaluated using single measurements by the new device compared to results of the standardized test method, and this evaluation must demonstrate a total error less than or equal to 6 percent.
(iv) Performance testing must demonstrate that there is little to no interference from common hemoglobin variants, including Hemoglobin C, Hemoglobin D, Hemoglobin E, Hemoglobin A2, and Hemoglobin S.
(3) When assay interference from Hemoglobin F or interference with other hemoglobin variants with low frequency in the population is observed, a warning statement must be placed in a black box and must appear in all labeling material for these devices describing the interference and any affected populations.