(114 days)
The HbA 1c (Hemoglobin A 1c) Advanced assay on the Beckman Coulter DxC700 AU Clinical Chemistry Analyzer, is intended for the quantitative determination of mmol/mol HbA1c (DCCT/NGSP) concentration in human venous whole blood. The determination of HbA1c is used as an aid in the diagnosis of diabetes mellitus, for the monitoring of long-term glucose control in individuals with diabetes mellitus and identifying patients who may be at risk for developing diabetes mellitus. For in vitro diagnostic use only.
The HbA1c Advanced reagent kit is in a liquid format and is ready to use. It contains four reagents HbA1c R1 and HbA1c R2, Total Hemoglobin R1 and Hemolyzing reagent R1. The HbA1c calibrator is supplied with the reagent, in a liquid, ready to use format and contains 5 x 2mL calibrator levels. The sample hemolysis is automated on the DxC700 AU Clinical Chemistry analyzer. Sample handling is performed as follows: 200 uL of hemolyzing reagent is aspirated from the Hemolyzing Reagent R1and dispensed into a cuvette. Tetradecyltrimethylammonium bromide (TTAB) in the hemolyzing reagent eliminates interference from leukocytes. 2 µL of whole blood sample is then aspirated from the patient sample and added to the hemolyzing reagent in the cuvette. This hemolyzed whole blood is then added to the THb assay cuvette and HbA1c assay cuvette as per the assay parameters. The concentrations of both HbA1c and Total Hemoglobin are determined. The HbA1c/Total Hemoglobin ratio is expressed either as mmol/mol (IFCC) or %HbA1c (DCCT/NGSP). Total Hemoglobin Reagent is used to measure total hemoglobin concentration by a colorimetric method. Change in absorbance is measured at 570/660 nm. HbA1c reagent is used to measure hemoglobin A1c concentration by a turbidimetric immunoinhibition method. In the reaction, hemoglobin A1c antibodies combine with HbA1c from the sample to form soluble antigen-antibody complexes. Polyhaptens from the reagent then bind with the excess antibodies and the resulting agglutinated complex is measured turbidimetrically. Change in absorbance is measured at 340/700 nm.
Here's an analysis of the acceptance criteria and study details for the HbA1c Advanced device, based on the provided document:
Acceptance Criteria and Device Performance
| Parameter | Acceptance Criteria | Reported Device Performance (NGSP Units) | Pass/Fail |
|---|---|---|---|
| Precision | |||
| Repeatability (Within-run) | CV ≤1.5% or SD ≤0.1% HbA1c | Human Whole Blood 1 (5.06%): 0.88% CV, 0.04 SD Human Whole Blood 2 (6.72%): 1.01% CV, 0.07 SD Human Whole Blood 3 (8.06%): 0.77% CV, 0.06 SD Human Whole Blood 4 (11.70%): 0.79% CV, 0.09 SD Spiked Human Whole Blood (14.02%): 0.74% CV, 0.10 SD Whole Blood Control 1 (5.32%): 1.19% CV, 0.06 SD Whole Blood Control 2 (9.88%): 0.77% CV, 0.08 SD | Pass |
| Total Precision | CV ≤2% or SD ≤0.13% HbA1c | Human Whole Blood 1 (5.06%): 1.63% CV, 0.08 SD Human Whole Blood 2 (6.72%): 1.64% CV, 0.11 SD Human Whole Blood 3 (8.06%): 1.57% CV, 0.13 SD Human Whole Blood 4 (11.70%): 1.26% CV, 0.15 SD Spiked Human Whole Blood (14.02%): 1.19% CV, 0.17 SD Whole Blood Control 1 (5.32%): 2.08% CV, 0.11 SD Whole Blood Control 2 (9.88%): 1.54% CV, 0.15 SD | Pass |
| Linearity (NGSP) | |||
| Linear Range | 4-15% HbA1c | 3.94% HbA1c to 15.37% HbA1c | Pass |
| Regression Parameters | Slope: 1.0 ± 0.05; Intercept: ≤ ± 0.5 % HbA1c; R: ≥ 0.990; N: ≥ 9 | Slope: 1.0 ± 0.05; Intercept: ≤ ± 0.5 % HbA1c; R: ≥ 0.990; N: ≥ 9 (All met) | Pass |
| Method Comparison (NGSP) | |||
| Slope (Weighted Deming) | 1.0 ± 0.05 | 0.990 (0.978; 1.002) | Pass |
| Intercept (Weighted Deming) | ≤ ± 0.5% HbA1c | 0.010 %HbA1c (-0.070; 0.089) %HbA1c | Pass |
| R (Weighted Deming) | ≥ 0.975 | 0.998 | Pass |
| Slope (Passing-Bablok) | 1.0 ± 0.5 | 0.980 (0.964; 0.992) | Pass |
| Intercept (Passing-Bablok) | ≤ ± 0.5% HbA1c | 0.090 %HbA1c (-0.006; 0.187) % HbA1c | Pass |
| R (Passing-Bablok) | ≥ 0.975 | 0.998 | Pass |
| Total Error | ≤6% | 5.0% HbA1c: 4.3% 6.5% HbA1c: 4.2% 8.0% HbA1c: 4.3% 12.0% HbA1c: 3.3% | Pass |
| Analytical Specificity | No Significant Interference (recovery within 7% of initial value) | Endogenous Interference: No significant interference up to stated concentrations for Conjugated Bilirubin (60 mg/dL), Unconjugated Bilirubin (60 mg/dL), Lipemia (500 mg/dL), Ascorbic Acid (300 mg/dL), RF (1000 IU/ml), Total Protein (21 g/dL), Glucose (2000 mg/dL). Drug Interference: No significant interference up to stated concentrations for numerous drugs (e.g., Glyburide 0.12 mg/dL, Salicylic Acid 4.76 mg/dL, Acetaminophen 26 mg/dL, etc.). Hemoglobin Derivative and Cross Reactants: No significant interference up to stated concentrations for Labile Hemoglobin (2000 mg/dL), Acetylated Hemoglobin (0.5 mg/mL), Carbamylated Hemoglobin (1.5 mg/mL), Glycated Albumin (5mg/mL), HbA0 (12 mg/mL), HbA1a + 1b (0.16 mg/mL). | Pass |
| Hemoglobin Variants (Bias) | No Significant Interference (recovery within 7% of reference value) | HbC: -2.57% bias (range -4.30% to -1.80%) at ~6.5% HbA1c; -3.19% bias (range -6.48% to 0.41%) at ~9.0% HbA1c. HbD: -0.77% bias (range -4.81% to 2.99%) at ~6.5% HbA1c; -1.22% bias (range -6.30% to -0.22%) at ~9.0% HbA1c. HbE: -1.12% bias (range -9.16% to 2.48%) at ~6.5% HbA1c; 0.47% bias (range -1.76% to 4.21%) at ~9.0% HbA1c. HbS: -1.18% bias (range -2.17% to 3.04%) at ~6.5% HbA1c; -1.04% bias (range -3.33% to 4.41%) at ~9.0% HbA1c. HbA2: 0.48% bias (range -1.92% to 5.60%) at ~6.5% HbA1c; 2.49% bias (range -0.98% to 3.60%) at ~9.0% HbA1c. HbF: Specimens containing >7% HbF may yield lower than expected HbA1c values (disclaimer). | Pass (with disclaimer for HbF) |
Study Details
-
Sample size used for the test set and the data provenance:
- Precision (EP05-A3): Four levels of HbA1c K2 EDTA human venous whole blood patient samples (approx. 5.0%, 6.5%, 8.0%, 12%, and 14%) and two whole blood controls. Samples were analyzed in duplicate, twice daily, over 20 working days on 3 different instrument lots. (n=2 for each sample/control, 2x daily, 20 days: 80 measurements per sample/control per instrument, total for all samples/controls/instruments: 8073 = 1680 individual measurements for precision)
- Linearity (EP06-A): High and low pools of human whole blood were used to create a linearity series to span the analytical range. The exact number of samples in the linearity series is implied to be at least 9 (N: ≥ 9 for regression parameters).
- Method Comparison (EP09-A3): 138 venous human frozen whole blood specimens (K2 EDTA anticoagulant type). These were patient samples. The provenance is not explicitly stated as country of origin, but they are referred to as "venous human frozen whole blood specimens." The study design is prospective in the sense that the samples were collected and then tested for the study; it is not explicitly called out as retrospective/prospective.
- Analytical Specificity (Interference - EP07): Two % HbA1c concentrations (approx. 6.5% and 8.0% HbA1c) using low and high pools prepared from human whole blood. Interfering substances were tested at a minimum of 5 levels each, with 10 replicates per level. The interference assessment was done on human venous whole blood K2 EDTA samples.
- Hemoglobin Variants: A minimum of 20 samples were tested for each variant (HbC, HbD, HbE, HbF, HbS, HbA2), totaling at least 120 samples. These are patient samples with identified hemoglobin variants.
-
Number of experts used to establish the ground truth for the test set and the qualifications of those experts:
- For the Method Comparison study, the ground truth was established by an "NGSP Secondary Reference Laboratory (SRL) using a test system (method X - HA8180V HPLC)." The specific number and qualifications of experts at the SRL are not detailed in this document.
- For Hemoglobin Variants, the reference methods used to establish the ground truth were "Trinity Biotech Hb9210 and Ultra2, Menarini HA8181V and TOSOH G8," which are demonstrated to be free from hemoglobin interference. Again, the number and qualifications of the operators of these reference methods are not specified.
-
Adjudication method (e.g. 2+1, 3+1, none) for the test set:
- The document does not describe an adjudication method involving multiple human readers for establishing ground truth. The ground truth for method comparison and hemoglobin variant studies relies on reference methods/laboratories.
-
If a multi-reader multi-case (MRMC) comparative effectiveness study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance:
- No MRMC comparative effectiveness study involving human readers or AI assistance was conducted or described, as this device is an in-vitro diagnostic (IVD) test system (HbA1c assay) and not an imaging AI device that would typically involve human reader interpretation.
-
If a standalone (i.e. algorithm only without human-in-the-loop performance) was done:
- Yes, the performance characteristics described are for the "HbA1c Advanced reagent on the DxC 700 AU Clinical Chemistry Analyzer" as a standalone device (algorithm only performance, in a laboratory setting). The entire submission details the analytical performance of the device itself.
-
The type of ground truth used (expert consensus, pathology, outcomes data, etc.):
- Precision: Internal validation against the device's own specifications.
- Linearity: Internal validation for demonstrating accurate measurement across specific concentration ranges.
- Method Comparison: Comparison to an NGSP Secondary Reference Laboratory (SRL) standardized method (HA8180V HPLC).
- Analytical Specificity (Interference): Comparison to baseline measurements of samples without interferents, assessed against a 7% recovery criteria.
- Hemoglobin Variants: Comparison to reference methods demonstrated to be free from hemoglobin interference (Trinity Biotech Hb9210 and Ultra2, Menarini HA8181V and TOSOH G8).
-
The sample size for the training set:
- This document describes the analytical validation of a biochemical assay on an automated analyzer. There is no "training set" in the context of machine learning, as the device is a reagent and instrument system, not an AI/ML algorithm that is trained on data. The device's performance is rigorously tested as described above.
-
How the ground truth for the training set was established:
- Not applicable, as this is not an AI/ML device with a training set. The assay's analytical parameters are established through chemical and immunochemical principles and optimized during development.
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Image /page/0/Picture/0 description: The image shows the logo of the U.S. Food and Drug Administration (FDA). The logo consists of two parts: the Department of Health and Human Services logo on the left and the FDA logo on the right. The FDA logo is a blue square with the letters "FDA" in white, followed by the words "U.S. Food & Drug Administration" in blue.
January 16, 2019
Beckman Coulter Ireland Inc. Catriona Hourigan, Regulatory Affairs Specialist Lismeehan O' Callaghan's Mills Co. Clare Ireland V94 PP63
Re: K182651
Trade/Device Name: HbA1c Advanced Regulation Number: 21 CFR 862.1373 Regulation Name: Hemoglobin A1c test system Regulatory Class: Class II Product Code: PDJ, LCP Dated: September 20, 2018 Received: September 24, 2018
Dear Catriona Hourigan:
We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.
If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.
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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 Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/CombinationProducts/GuidanceRegulatoryInformation/ucm597488.html; good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.
Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm.
For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/MedicalDevices/DeviceRegulationandGuidance/) and CDRH Learn (http://www.fda.gov/Training/CDRHLearn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (http://www.fda.gov/DICE) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).
Sincerely,
Kellie B. Kelm -S
Courtney H. Lias, Ph.D. for 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) K182651
Device Name HbA1c Advanced
Indications for Use (Describe)
The HbA 1c (Hemoglobin A 1c) Advanced assay on the Beckman Coulter DxC700 AU Clinical Chemistry Analyzer, is intended for the quantitative determination of mmol/mol HbA1c (DCCT/NGSP) concentration in human venous whole blood. The determination of HbA1c is used as an aid in the diagnosis of diabetes mellitus, for the monitoring of long-term glucose control in individuals with diabetes mellitus and identifying patients who may be at risk for developing diabetes mellitus. For in vitro diagnostic use only.
Type of Use (Select one or both, as applicable)
| ☑ Prescription Use (Part 21 CFR 801 Subpart D) |
|---|
| ☐ Over-The-Counter Use (21 CFR 801 Subpart C) |
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510(k) Summary
HbA1c Advanced
1.0 Submitted By:
Catriona Hourigan Regulatory Affairs Specialist Beckman Coulter Ireland Inc., Lismeehan, O' Callaghan's Mills, Co. Clare, Ireland. Phone: +353-65-683-1417 FAX: +353-65-683-1122 email: chourigan@beckman.com
2.0 Date Submitted:
15th January 2019
3.0 Device Name(s):
Proprietary Name: HbA1c Advanced K182651
4.0 Predicate Device:
Table 1: Predicate Details
| Candidate(s) | Predicate | Manufacturer | DocketNumber |
|---|---|---|---|
| HbA1c Advanced | D-100™ HbA1c,D-100™HbA1cCalibrator Pack | Bio-RadLaboratories, Inc; | K151321 |
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5.0 Recommended Reagent Classification: Name: Hemoglobin A1c Test System Requlation: 862.1373 Class: II
Name: Assay Glycosylated Hemoglobin Regulation: 864.7470 Class: II Panel: Hematology Product Code: LCP
Panel: Clinical Chemistry Product Code: PDJ
6.0 Description:
The HbA1c Advanced reagent kit is in a liquid format and is ready to use. It contains four reagents HbA1c R1 and HbA1c R2, Total Hemoglobin R1 and Hemolyzing reagent R1. The HbA1c calibrator is supplied with the reagent, in a liquid, ready to use format and contains 5 x 2mL calibrator levels. The sample hemolysis is automated on the DxC700 AU Clinical Chemistry analyzer. Sample handling is performed as follows: 200 uL of hemolyzing reagent is aspirated from the Hemolyzing Reagent R1and dispensed into a cuvette. Tetradecyltrimethylammonium bromide (TTAB) in the hemolyzing reagent eliminates interference from leukocytes. 2 µL of whole blood sample is then aspirated from the patient sample and added to the hemolyzing reagent in the cuvette. This hemolyzed whole blood is then added to the THb assay cuvette and HbA1c assay cuvette as per the assay parameters.
The concentrations of both HbA1c and Total Hemoglobin are determined. The HbA1c/Total Hemoglobin ratio is expressed either as mmol/mol (IFCC) or %HbA1c (DCCT/NGSP).
Total Hemoglobin Reagent is used to measure total hemoglobin concentration by a colorimetric method. Change in absorbance is measured at 570/660 nm.
HbA1c reagent is used to measure hemoglobin A1c concentration by a turbidimetric immunoinhibition method. In the reaction, hemoglobin A1c antibodies combine with HbA1c from the sample to form soluble antigen-antibody complexes. Polyhaptens from the reagent then bind with the excess antibodies and the resulting agglutinated complex is measured turbidimetrically. Change in absorbance is measured at 340/700 nm.
The calibrator is manufactured from human material; therefore should be handled as though capable of transmitting infectious disease. Each donor used in the preparation of this material was tested by the United States Food and Drug Administration (FDA), approved method and found to be negative for the presence of the antibodies for HIV-1/2, HCV, Hepatitis B surface antigen and was determined to not be repeatedly reactive.
Because no test method can offer complete assurance that HIV-1/2, HCV, hepatitis B virus
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or other infectious agents are absent from biological materials, this product should be handled at the Biosafety Level 2 as recommended for any infectious human serum or blood specimen in the Centers for Disease Control and Prevention/National Institutes of Health manual, Biosafety in Microbiological and Biomedical Laboratories.
7.0 Intended Use:
HBA1c Advanced
The HbA1c Advanced assay on the Beckman Coulter DxC 700 AU Clinical Chemistry Analyzer, is intended for the quantitative determination of mmol/mol HbA1c (IFCC) and % HbA1c (DCCT/NGSP) concentration in human venous whole blood. The determination of HbA1c is used as an aid in diagnosis of diabetes mellitus, for the monitoring of long-term blood glucose control in individuals with diabetes mellitus and identifying patients who may be at risk for developing diabetes mellitus. For In vitro diagnostic use only.
8.0 Comparison to the Predicate(s):
| Test System | HbA1c Predicate | Proposed New System |
|---|---|---|
| Proprietary andEstablished Names | D-100™ HbA1c,D-100™ HbA1c CalibratorPack | HbA1c Advanced |
| Similarities | ||
| Intended use | The D-100™ HbA1c test isintended for the quantitativedetermination ofhemoglobin A1c (IFCCmmol/mol and NGSP %) inhuman whole blood usingion-exchange highperformance liquidchromatography (HPLC) onthe D-100 Hemoglobintesting system.Hemoglobin A1cmeasurements are used asan aid in the diagnosis ofdiabetes mellitus, as an aidto identify patients who maybe at risk for developingdiabetes mellitus, and forthe monitoring of long-term | The HbA1c (Hemoglobin A1c)Advanced assay on theBeckman Coulter DxC 700 AUClinical Chemistry Analyzer, isintended for the quantitativedetermination of mmol/molHbA1c (IFCC) or % HbA1c(DCCT/NGSP) concentration inhuman venous whole blood. Thedetermination of HbA1c is usedas an aid in the diagnosis ofdiabetes mellitus, for themonitoring of long-term bloodglucose control in individualswith diabetes mellitus andidentifying patients who may beat risk for developing diabetesmellitus. For in vitro diagnosticuse only. |
| blood glucose control inindividuals with diabetesmellitus.Calibrators:The D-100™ HbA1cCalibrator pack is for thecalibration of the D-100Hemoglobin Testing Systemused for the quantitativedetermination ofhemoglobin A1c (HbA1c) inhuman whole blood. | ||
| Reporting Units | HbA1c results are providedto the customers using twodifferent units:NGSP equivalent units (%)and IFCC equivalent units(mmol/mol). | Same |
| Sample Types | Whole bloodK2-EDTAK3-EDTALithium HeparinSodium Heparin | Whole bloodK2-EDTAK3-EDTALi-HeparinNa-Heparin |
| Reagent Stability | Unopened2° - 8°C until expiration date | Unopened2° - 8°C until expiration date |
| Reference Range | Literature reference:Diabetic ≥6.5 NGSP %, ≥48IFCC mmol/mol;Pre-Diabetic5.7 - 6.4 NGSP%, IFCC 39- 47 mmol/mol;Non--Diabetic<5.7NGSP%, <39 IFCCmmol/mol | Same |
| Traceability | Traceable to the DiabetesControl and Complicationstrial (DCCT) reference | Same |
| method and IFCC. Certifiedvia the NationalGlycohemoglobinStandardization Program(NGSP). | ||
| Sensitivity | Not stated | Not stated |
| Specificity(Interference -Hb Variants) | Significant interference wasdefined as ≥ 7% change inHbA1c value in thepresence of the hemoglobinvariant relative to control. | No Significant interference (NSI)is recovery within 7% of thereference value from Hbvariants C, S, D, E, A2. |
| HbC, HbD, HbE, HbSHbA2, HbF | Hemoglobin F interference shallbe displayed as a disclaimer onpackage insert. | |
| NSI is observed from thefollowing endogenoussubstances up to the statedconcentrations (Significantinterference is defined as≥7% change in HbA1cvalue of the mean test valuerelative to mean of thereference samples): | The criteria for NSI is recoverywithin 7% of the initial value(sample containing nointerferent). Similar for: | |
| Specificity(Interferences -Endogenous) | Conjugated Bilirubin:NSI up to 60 mg/dL Unconjugated Bilirubin:NSI up 60 mg/dL | Conjugated Bilirubin: NSIup to 60 mg/dL Unconjugated Bilirubin:NSI up to 60 mg/dL |
| Glucose: NSI up 2000mg/dL | Glucose: NSI up to 2000mg/dL | |
| Total Protein: NSI up to21 g/dL | Total Protein: NSI up to21 g/dL | |
| Specificity(Interference -Hb Derivatives andCross Reactants) | Significant interferencedefined as % recovery of±7% change in % HbA1cvalue from the control Acetylated Hb: up to 50mg/dL | NSI is recovery within 7% of theinitial value (sample containingno interferent). Acetylated Hb: NSI up to0.5 mg/mL |
| Specificity(Interference - Drug) | Significant interference wasdefined as a ≥7% change in%HbA1c value from thecontrol.Acetylcysteine: 166 mg/dLAmpicillin-Na: 1000 mg/dLAcetylsalicylic Acid: 1000mg/dLAscorbic Acid: 300 mg/dLCefoxitin: 2500 mg/dLLevodopa: 20 mg/dLMethyldopa: 20 mg/dLMetronidazole: 200 mg/dLDoxycyclin: 50 mg/dLCyclosporine: 5 mg/LTheophylline: 100 mg/LIbuprofen: 500 mg/LTheophylline: 100mg/L | The criteria for no significantinterference (NSI) is recoverywithin 7% of the initial value(Sample containing nointerferent):Acetylcystein:166 mg/dLAmpicillin-Na: 1000 mg/dLAcetylsalicylic Acid: 1000mg/dLAscorbic Acid: 300 mg/dLCefoxitin: 2500 mg/dLLevodopa: 20 mg/dLMethyldopa: 20 mg/dLMetronidazole: 200 mg/dLDoxycyclin: 50 mg/dLCyclosporine: 0.5 mg/dLTheophylline: 10 mg/dLIbuprofen: 50 mg/dL |
| Total Allowable Error | ≤6% | ≤6% |
Table 2: Similarities between the Predicate Device and the Proposed Device:
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Table 3: Differences between the Predicate Device and the Proposed Device:
| Feature | Predicate | Proposed New System |
|---|---|---|
| Proprietary andEstablished Names | D-100™ HbA1c,D-100™ HbA1c Calibrator Pack | HbA1c Advanced |
| Instrument Platform | D-100™ Hemoglobin TestingSystem | DxC 700 AU ClinicalChemistry Analyzer |
| Control | Commercially available Bio-Rad Lyphocheck Diabetescontrol materials | 2 Levels of control material |
| Operating Principle | Ion-exchange Quantitativehigh performance liquidchromatography (HPLC) | Quantitative turbidimetricinhibition immunoassay |
| Sample Types | Capillary bloodAcceptable anticoagulants arePotassium Oxalate/ Sodium FluorideSodium Citrate | Not Applicable for HbA1c Advanced |
| Analytical Range(Linearity) | HbA1c:3.5 to 20% HbA1c (NGSP)15 - 195 mmol/mol (IFCC) | HbA1c:4.0 - 15% HbA1c (NGSP)20 - 140 mmol/mol (IFCC) |
| Specificity(Interferences -Endogenous) | Significant interference defined as a ±7% change in % HbA1c value from the controlLipemia (Intralipid): NSI up to 6000 mg/dL Rheumatoid Factor: NSI up to 750 IU/mL | NSI is recovery within 7% of the initial value (sample containing no interferent).Lipemia: NSI up to 500 mg/dLRheumatoid Factor (RF): NSI up to 1000 IU/mL |
| Specificity(Interference -Hb Derivatives andCross Reactants) | Significant interference defined as % recovery of ±7% change in % HbA1c value from the control Carbamylated Hb: up to 5% Labile Hb: up to 1200 mg/dL of glucose does not interfere with this assay | NSI is recovery within 7% of the initial value (sample containing no interferent). Carbamylated Hb: NSI up to 1.5 mg/mL Labile Hb; NSI up to 2000 mg/dL Glycated Albumin: NSI up to 5 mg/mL HbA0: NSI up to 12 mg/mL |
| HbA1a+b: NSI up to 0.16 mg/mL | ||
| Drug Interference | Significant interference was defined as a ±7% change in %HbA1c value from the control. Acetaminophen: 200 mg/L Heparin: 5000 U/L Rifampicin: 64 mg/L Phenylbutazone – 400 mg/L | NSI is recovery within 7% of the initial value (sample containing no interferent): Acetaminophen: 26 mg/dL Heparin: 5500 IU/L Rifampicin: 8 mg/dL Phenylbutazone: 53.5 mg/dL Glyburide: 0.12 mg/dL Salicylic Acid: 4.76 mg/dL Sitagliptin: 0.2 mg/dL Rosiglitazone: 0.33 mg/dL Metformin: 5 mg/dL Calcium Dobsilate: 20mg/dL Acarbose: 0.05 mg/dL |
| Specificity(Interference -Hb Variants) | Significant interference was defined as ≥ 7% change in HbA1c value in the presence of the hemoglobin variant relative to control.HbF concentrations up to 30% do not interfere with the test. | No Significant interference is recovery within 7% of the reference value (value assigned in Secondary Reference Laboratory).Samples containing ≥7% HbF may result in lower than expected HbA1c results |
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9.0 Performance Characteristics – Analytical Performance:
Precision
Precision of the HbA1c Advanced reagent on the DxC 700 AU was evaluated based on CLSI guideline EP05-A3: "Evaluation of Precision Performance of Quantitative Measurement Methods; Approved Guideline - Third Edition". Studies were carried out on 3 lots of HbA1c Advanced by testing four levels of HbA1c K2 EDTA human venous whole blood patient samples at HbA1c concentrations of approximately 5.0%, 6.5%, 8.0%, 12% and 14%. The experimental design used duplicate sample analysis, twice daily, over the course of twenty working days (n=2) in random order. NGSP results are shown in Tables 4 - 7. IFCC results are shown in Tables 8 - 11.
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| Sample/ Controldetails | Mean %HbA1c | Repeatability(Within-run) | Specification | Between-Run | Between-Day | Between-Lot | TotalPrecision | Specification | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Concentration | %CV | SD | CV ≤1.5% or SD≤0.1% HbA1c | %CV | SD | %CV | SD | %CV | SD | %CV | SD | CV <2% orSD ≤0.13%HbA1c | |
| Human WholeBlood 1 | 5.06 | 0.88 | 0.04 | Pass | 0.80 | 0.04 | 0.64 | 0.03 | 0.92 | 0.05 | 1.63 | 0.08 | Pass |
| Human WholeBlood 2 | 6.72 | 1.01 | 0.07 | Pass | 0.60 | 0.04 | 0.65 | 0.04 | 0.93 | 0.06 | 1.64 | 0.11 | Pass |
| Human WholeBlood 3 | 8.06 | 0.77 | 0.06 | Pass | 0.47 | 0.04 | 0.73 | 0.06 | 1.06 | 0.09 | 1.57 | 0.13 | Pass |
| Human WholeBlood 4 | 11.70 | 0.79 | 0.09 | Pass | 0.48 | 0.06 | 0.84 | 0.10 | 0.19 | 0.02 | 1.26 | 0.15 | Pass |
| Spiked HumanWhole Blood | 14.02 | 0.74 | 0.10 | Pass | 0.49 | 0.07 | 0.72 | 0.10 | 0.32 | 0.04 | 1.19 | 0.17 | Pass |
| Whole BloodControl 1 | 5.32 | 1.19 | 0.06 | Pass | 0.84 | 0.04 | 0.66 | 0.03 | 1.34 | 0.07 | 2.08 | 0.11 | Pass |
| Whole BloodControl 2 | 9.88 | 0.77 | 0.08 | Pass | 0.48 | 0.05 | 0.65 | 0.06 | 1.07 | 0.11 | 1.54 | 0.15 | Pass |
Table 4: DxC 700 AU: Instrument 1: NGSP units: Summary of 20 Day Precision Performance
Table 5: DxC 700 AU Instrument 2: NGSP units: Summary of 20 Day Precision Performance
| Sample/ Controldetails | Mean %HbA1cConcentration | Repeatability(Within-run) | Specification | Between-Run | Between-Day | Between-Lot | TotalPrecision | Specification | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | ||||
| Human WholeBlood 1 | 5.03 | 1.15 | 0.06 | Pass | 0.37 | 0.02 | 0.57 | 0.03 | 1.02 | 0.05 | 1.68 | 0.08 | Pass |
| Human WholeBlood 2 | 6.69 | 1.14 | 0.08 | Pass | 0.20 | 0.01 | 0.50 | 0.03 | 0.83 | 0.06 | 1.51 | 0.10 | Pass |
| Human WholeBlood 3 | 8.05 | 0.86 | 0.07 | Pass | 0.48 | 0.04 | 0.49 | 0.04 | 0.70 | 0.06 | 1.31 | 0.11 | Pass |
| Human WholeBlood 4 | 11.69 | 0.71 | 0.08 | Pass | 0.33 | 0.04 | 0.61 | 0.07 | 0.56 | 0.07 | 1.14 | 0.13 | Pass |
| Spiked HumanWhole Blood | 14.04 | 0.75 | 0.11 | Pass | 0.58 | 0.08 | 0.34 | 0.05 | 0.69 | 0.10 | 1.22 | 0.17 | Pass |
| Whole BloodControl 1 | 5.28 | 1.23 | 0.06 | Pass | 0.41 | 0.02 | 0.75 | 0.04 | 1.32 | 0.07 | 1.99 | 0.11 | Pass |
| Whole BloodControl 2 | 9.88 | 0.77 | 0.08 | Pass | 0.42 | 0.04 | 0.54 | 0.05 | 0.95 | 0.09 | 1.40 | 0.14 | Pass |
{13}------------------------------------------------
| Sample/ Controldetails | Repeatability(Within-run) | Specification | Between-Run | Between-Day | Between-Lot | TotalPrecision | Specification | ||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Mean %HbA1cConcentration | %CV | SD | CV<1.5% or SD<0.1% HbA1c | %CV | SD | %CV | SD | %CV | SD | %CV | SD | CV <2% or SD<0.13% HbA1c | |
| Human WholeBlood 1 | 5.03 | 0.83 | 0.04 | Pass | 0.45 | 0.02 | 0.70 | 0.04 | 1.02 | 0.05 | 1.55 | 0.08 | Pass |
| Human WholeBlood 2 | 6.66 | 0.82 | 0.06 | Pass | 0.31 | 0.02 | 0.48 | 0.03 | 1.31 | 0.09 | 1.65 | 0.11 | Pass |
| Human wholeBlood 3 | 7.98 | 0.73 | 0.06 | Pass | 0.58 | 0.05 | 0.47 | 0.04 | 1.68 | 0.14 | 1.98 | 0.16 | Pass |
| Human WholeBlood 4 | 11.68 | 0.71 | 0.08 | Pass | 0.21 | 0.03 | 0.67 | 0.08 | 0.54 | 0.06 | 1.14 | 0.13 | Pass |
| Spiked HumanWhole Blood | 13.99 | 0.66 | 0.09 | Pass | 0.44 | 0.06 | 0.56 | 0.08 | 0.66 | 0.09 | 1.18 | 0.17 | Pass |
| Whole BloodControl 1 | 5.25 | 0.93 | 0.05 | Pass | 0.97 | 0.05 | 0.60 | 0.03 | 1.37 | 0.07 | 2.01 | 0.11 | Pass |
| Whole BloodControl 2 | 9.72 | 0.65 | 0.06 | Pass | 0.36 | 0.03 | 0.65 | 0.06 | 1.45 | 0.14 | 1.75 | 0.17 | Pass |
Table 6: DxC 700 AU Instrument 3: NGSP units: Summary of 20 Day Precision Performance
{14}------------------------------------------------
| Sample/ Controldetails | Mean %HbA1cConcentration | Repeatability(Within-run) | Specification | Between-Run | Between-Day | Between-Lot | BetweenInstrument | TotalPrecision | Specification | ||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| %CV | SD | CV<=1.5% or SD<=0.1% HbA1c | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | CV <=2% orSD <=0.13%HbA1c | ||
| Human WholeBlood 1 | 5.07 | 0.96 | 0.05 | Pass | 0.57 | 0.03 | 0.64 | 0.03 | 0.99 | 0.05 | 0.74 | 0.04 | 1.78 | 0.09 | Pass |
| Human WholeBlood 2 | 6.72 | 1.00 | 0.07 | Pass | 0.41 | 0.03 | 0.55 | 0.04 | 1.05 | 0.07 | 0.56 | 0.04 | 1.69 | 0.11 | Pass |
| Human WholeBlood 3 | 8.07 | 0.79 | 0.06 | Pass | 0.51 | 0.04 | 0.58 | 0.05 | 1.22 | 0.10 | 0.57 | 0.05 | 1.74 | 0.14 | Pass |
| Human WholeBlood 4 | 11.71 | 0.74 | 0.09 | Pass | 0.36 | 0.04 | 0.71 | 0.08 | 0.46 | 0.05 | 0.32 | 0.04 | 1.22 | 0.14 | Pass |
| Spiked HumanWhole Blood | 14.03 | 0.72 | 0.10 | Pass | 0.51 | 0.07 | 0.56 | 0.08 | 0.58 | 0.08 | 0.12 | 0.02 | 1.20 | 0.17 | Pass |
| Whole BloodControl 1 | 5.29 | 1.12 | 0.06 | Pass | 0.78 | 0.04 | 0.67 | 0.04 | 1.34 | 0.07 | 0.50 | 0.03 | 2.09 | 0.11 | Pass |
| Whole BloodControl 2 | 9.84 | 0.73 | 0.07 | Pass | 0.43 | 0.04 | 0.61 | 0.06 | 1.17 | 0.12 | 0.70 | 0.07 | 1.72 | 0.17 | Pass |
Table 7: Instruments Combined (Instrument 1, 2, 3): NGSP units: Summary of 20 Day Precision Performance
{15}------------------------------------------------
| Sample/ Controldetails | Mean(mmol/mol) | Repeatability(Within-run) | Specification | Between-Run | Between-Day | Between-Lot | TotalPrecision | Specification | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| HbA1cConcentration | %CV | SD | CV <2.3% or SD<1.1 mmol/mol | %CV | SD | %CV | SD | %CV | SD | %CV | SD | CV ≤2.9% or SD<1.4 mmol/mol | |
| Human WholeBlood 1 | 31.84 | 1.51 | 0.48 | Pass | 1.43 | 0.46 | 1.08 | 0.34 | 1.60 | 0.51 | 2.84 | 0.90 | Pass |
| Human WholeBlood 2 | 49.92 | 1.49 | 0.75 | Pass | 0.90 | 0.45 | 0.97 | 0.48 | 1.36 | 0.68 | 2.41 | 1.21 | Pass |
| Human WholeBlood 3 | 64.65 | 1.06 | 0.68 | Pass | 0.61 | 0.40 | 1.01 | 0.65 | 1.43 | 0.93 | 2.14 | 1.38 | Pass |
| Human WholeBlood 4 | 104.38 | 1.31 | 1.37 | Pass | 0.00 | 0.00 | 1.03 | 1.07 | 0.23 | 0.24 | 1.68 | 1.76 | Pass |
| Spiked HumanWhole Blood | 129.76 | 0.87 | 1.13 | Pass | 0.57 | 0.74 | 0.86 | 1.11 | 0.37 | 0.49 | 1.40 | 1.82 | Pass |
| Whole BloodControl 1 | 34.65 | 2.01 | 0.70 | Pass | 1.28 | 0.44 | 1.18 | 0.41 | 2.22 | 0.77 | 3.46 | 1.20 | Pass |
| Whole BloodControl 2 | 84.53 | 0.98 | 0.83 | Pass | 0.61 | 0.51 | 0.82 | 0.69 | 1.36 | 1.15 | 1.96 | 1.66 | Pass |
Table 8: DxC 700 AU Instrument 1: IFCC units: Summary of 20 Day Precision Performance
{16}------------------------------------------------
| Sample/Control details | Mean(mmol/mol)HbA1cConcentration | Repeatability(Within-run) | Specification | Between-Run | Between-Day | Between-Lot | TotalPrecision | Specification | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| %CV | SD | CV ≤2.3%or SD ≤1.1mmol/mol | %CV | SD | %CV | SD | %CV | SD | %CV | SD | CV ≤2.9%or SD ≤1.4mmol/mol | ||
| Human WholeBlood 1 | 31.51 | 2.01 | 0.63 | Pass | 1.19 | 0.37 | 0.77 | 0.24 | 1.81 | 0.57 | 3.05 | 0.96 | Pass |
| Human WholeBlood 2 | 49.59 | 1.63 | 0.81 | Pass | 0.42 | 0.21 | 0.74 | 0.37 | 1.18 | 0.59 | 2.18 | 1.08 | Pass |
| Human WholeBlood 3 | 64.48 | 1.18 | 0.76 | Pass | 0.66 | 0.43 | 0.67 | 0.43 | 0.96 | 0.62 | 1.79 | 1.15 | Pass |
| Human WholeBlood 4 | 104.29 | 0.88 | 0.91 | Pass | 0.38 | 0.40 | 0.75 | 0.78 | 0.70 | 0.73 | 1.40 | 1.46 | Pass |
| Spiked HumanWhole Blood | 129.92 | 0.88 | 1.15 | Pass | 0.66 | 0.85 | 0.43 | 0.56 | 0.82 | 1.06 | 1.44 | 1.87 | Pass |
| Whole BloodControl 1 | 34.24 | 2.09 | 0.72 | Pass | 0.74 | 0.25 | 1.23 | 0.42 | 2.23 | 0.76 | 3.38 | 1.16 | Pass |
| Whole BloodControl 2 | 84.52 | 0.98 | 0.83 | Pass | 0.54 | 0.46 | 0.69 | 0.58 | 1.22 | 1.03 | 1.80 | 1.52 | Pass |
Table 9: DxC 700 AU Instrument 2: IFCC units: Summary of 20 Day Precision Performance
{17}------------------------------------------------
| Sample/Control details | Mean(mmol/mol) | Repeatability(Within-run) | Specification | Between-Run | Between-Day | Between-Lot | TotalPrecision | Specifications | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| HbA1cConcentration | %CV | SD | CV <2.3%or SD <1.1mmol/mol | %CV | SD | %CV | SD | %CV | SD | %CV | SD | CV ≤2.9%or SD <1.4mmol/mol | |
| Human WholeBlood 1 | 31.46 | 1.41 | 0.46 | Pass | 1.18 | 0.38 | 1.31 | 0.42 | 1.78 | 0.58 | 2.88 | 0.93 | Pass |
| Human WholeBlood 2 | 49.27 | 1.21 | 0.61 | Pass | 0.46 | 0.23 | 0.70 | 0.35 | 1.93 | 0.97 | 2.42 | 1.22 | Pass |
| Human WholeBlood 3 | 63.68 | 1.00 | 0.65 | Pass | 0.79 | 0.52 | 0.63 | 0.41 | 2.29 | 1.49 | 2.69 | 1.75 | Pass |
| Human WholeBlood 4 | 104.18 | 0.87 | 0.92 | Pass | 0.25 | 0.26 | 0.81 | 0.85 | 0.66 | 0.70 | 1.39 | 1.46 | Pass |
| Spiked HumanWhole Blood | 129.37 | 0.79 | 1.02 | Pass | 0.52 | 0.67 | 0.66 | 0.86 | 0.87 | 1.01 | 1.39 | 1.81 | Pass |
| Whole BloodControl 1 | 33.83 | 1.58 | 0.54 | Pass | 1.61 | 0.55 | 1.02 | 0.35 | 2.32 | 0.79 | 3.40 | 1.16 | Pass |
| Whole BloodControl 2 | 82.77 | 0.83 | 0.69 | Pass | 0.46 | 0.38 | 0.83 | 0.69 | 1.86 | 1.55 | 2.24 | 1.87 | Pass |
Table 10: DxC 700 AU Instrument 3: IFCC units: Summary of 20 Day Precision Performance
{18}------------------------------------------------
| Sample/Control details | Mean(mmol/mol)HbA1cConcen-tration | Repeatability(Within-run) | Specification | Between-Run | Between-Day | Between-Lot | BetweenInstrument | TotalPrecision | Specification | ||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| % CV | SD | CV≤2.3% or SD≤1.1mmol/mol | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | CV ≤2.9% orSD ≤1.4mmol/mol | ||
| Human WholeBlood 1 | 31.91 | 1.66 | 0.53 | Pass | 1.27 | 0.41 | 1.08 | 0.34 | 1.73 | 0.55 | 1.31 | 0.42 | 3.20 | 1.02 | Pass |
| Human WholeBlood 2 | 49.95 | 1.45 | 0.72 | Pass | 0.63 | 0.31 | 0.81 | 0.41 | 1.53 | 0.76 | 0.82 | 0.41 | 2.49 | 1.24 | Pass |
| Human WholeBlood 3 | 64.75 | 1.08 | 0.70 | Pass | 0.69 | 0.45 | 0.79 | 0.51 | 1.66 | 1.07 | 0.77 | 0.50 | 2.37 | 1.53 | Pass |
| Human WholeBlood 4 | 104.51 | 1.04 | 1.09 | Pass | 0.23 | 0.24 | 0.87 | 0.91 | 0.57 | 0.60 | 0.40 | 0.42 | 1.54 | 1.61 | Pass |
| Spiked HumanWhole Blood | 129.84 | 0.85 | 1.10 | Pass | 0.58 | 0.76 | 0.67 | 0.87 | 0.69 | 0.89 | 0.14 | 0.19 | 1.42 | 1.84 | Pass |
| Whole BloodControl 1 | 34.32 | 1.91 | 0.66 | Pass | 1.26 | 0.43 | 1.15 | 0.39 | 2.26 | 0.78 | 0.87 | 0.30 | 3.52 | 1.21 | Pass |
| Whole BloodControl 2 | 84.10 | 0.93 | 0.78 | Pass | 0.54 | 0.46 | 0.78 | 0.66 | 1.50 | 1.26 | 0.90 | 0.76 | 2.20 | 1.85 | Pass |
Table 10: Instruments Combined (Instrument 1, 2, 3): IFCC units: Summary of 20 Day Precision Performance
{19}------------------------------------------------
Linearity:
Dynamic Range/ Analytical Measuring Range studies were designed using CLSI Guideline EP06-A: "Evaluation of the Linearity of Quantitative Measurement Procedures: A Statistical Approach; Approved Guideline". High and low pools were prepared as per Table 1. A linearity series was prepared by inter-diluting the high and low pools to achieve a % HbA1c concentration spanning the required analytical range. The samples were mixed together in varying ratios.
Data was analyzed using Polynomial regression analysis (for first, second and third order polynomials) to determine the statistical significance of non-linearity. Where data generated was non-linear, the best fitting polynomial was applied to the data and difference between this best fitting, non-linear polynomial and the linear polynomial was calculated. Regression parameters for slope, intercept and R value were calculated.
The results of the study provides data to support the dynamic range/analytical measuring range claim for HbA1c Advanced reagent 4 – 15% (NGSP) HbA1c and 20 – 140 mmol/mol HbA1c (IFCC).
| Reagent | AcceptanceCriteria | Results | Pass/Fail | AcceptanceCriteria | Pass/Fail | |
|---|---|---|---|---|---|---|
| LinearRange | LinearFrom | LinearTo | RegressionParameters | |||
| HbA1cAdvanced | 4-15% HbA1c | 3.94%HbA1c | 15.37%HbA1c | Pass | Slope: 1.0 ± 0.05Intercept: ≤ ± 0.5 % HbA1cR: ≥ 0.990N: ≥ 9 | Pass |
| HbA1cAdvanced | 20 – 140mmol/mol | 19.51mmol/mol | 144.49mmol/mol | Pass | Slope: 1.0 ± 0.05Intercept: ≤ ± 5.5 mmol/LR: ≥ 0.990N: ≥ 9 | Pass |
Table 12: Linearity Results Summary
{20}------------------------------------------------
Method Comparison
Method comparison and bias estimation experiments were designed using CLSI Guideline EP09-A3; "Measurement Procedure Comparison and Bias Estimation using Patient Samples; Approved Guideline." The patient correlation studies were performed using a standardized test method in an NGSP Secondary Reference Laboratory (SRL) using a test system (method X - HA8180V HPLC) with an analytical measuring range of 4-15% HbA1c (NGSP) and the proposed new Beckman Coulter HbA1c test system (method Y - HbA1c Advanced, 1 lot) with a proposed measuring range of 4-15 % HbA1c (NGSP) on a DxC 700 AU analyzer. Over a 120 (duplicates) of venous human frozen whole blood specimens (K2 EDTA anticoagulant type) with HbA1c concentrations were tested using on the DxC 700 AU Analyzer.
| HemoglobinA1c Level | No. ofSamples(SRL) | % Samplestested (SRL) | No. of Samplestested (Beckman) | % Samplestested (Beckman) |
|---|---|---|---|---|
| ≤5% | 6 | 4.35 | 6 | 4.35 |
| 5 – 6% | 18 | 13.04 | 22 | 15.94 |
| 6 – 6.5% | 32 | 23.19 | 32 | 23.19 |
| 6.5 – 7% | 35 | 25.36 | 35 | 25.36 |
| 7 – 8% | 25 | 18.12 | 23 | 16.67 |
| 8 – 9% | 12 | 8.7 | 10 | 7.25 |
| > 9% | 10 | 7.25 | 10 | 7.25 |
| Total Samples | 138 | 100% | 138 | 100% |
Table 13: Sample Distribution
Weighted Deming's and Passing-Bablok regression analysis was performed for the HbA1c Advancedmethod versus an NGSP SRL standardized method, results are summarized below in tables 14 and 15.
{21}------------------------------------------------
Table 14: Summary of Method Comparison for HbA1c Advanced results and specifications (Weighted Deming analysis) NGSP Units.
| Sample Range: | Specifications | Results(95%CI Low; High) | PassFail |
|---|---|---|---|
| Method X:4.7 - 14.2 % HbA1c | Slope: 1.0 ± 0.05 | 0.990(0.978; 1.002) | Pass |
| Method Y:4.6 – 14.3 % HbA1c | Intercept: ≤ ± 0.5% HbA1c | 0.010 %HbA1c(-0.070; 0.089) %HbA1c | Pass |
| R: ≥ 0.975 | 0.998 | Pass | |
| N: ≥ 120 | 138 | Pass |
Table 15: Summary of Method Comparison for HbA1c Advanced results and specifications (Passing-Bablok analysis) NGSP Units.
| Sample Range: | Specifications | Results(95% CI Low; High) | Pass / Fail |
|---|---|---|---|
| Method X:4.7 - 14.2 % HbA1c | Slope: 1.0 ± 0.5 | 0.980(0.964;0.992) | Pass |
| Method Y:4.6 - 14.3 % HbA1c | Intercept: ≤ ± 0.5%HbA1cR: ≥ 0.975N: ≥ 120 | 0.090 %HbA1c(-0.006; 0.187) % HbA1c0.998138 | PassPassPass |
Total Error
Total error was evaluated using the results of the bias estimation (%Bias) from single measurements conducted of the new device compared to results of the standardised test method (method comparison) and precision estimates from the precision study. Total Error (TE) at four concentrations (5.0%, 6.5%, 8.0%, and 12.0%) was calculated as follows: "%TE = |(%Bias)] + 1.96*%CV*(1+%Bias/100)". The results are presented in Table 5.
Table 16: Total Error Estimation
| %HbA1cDecision Level | % Bias | %CV | %TE | AcceptanceCriteria:≤6% |
|---|---|---|---|---|
| 5.0 | -0.80 | 1.78 | 4.3 | Pass |
| 6.5 | -0.85 | 1.69 | 4.2 | Pass |
| 8.0 | -0.88 | 1.74 | 4.3 | Pass |
| 12.0 | -0.92 | 1.22 | 3.3 | Pass |
{22}------------------------------------------------
Analytical Specificity
Interference (Analytical Specificity) studies were designed based on CLSI Guideline EP07 Third Edition: "Interference Testing in Clinical Chemistry; Approved Guideline" and used to assess common or known substances which could interfere with the HbA1c Advanced assay.
Endogenous Interference
The interfering substances analyzed were tested at two % HbA1c concentrations; approximately 6.5% HbA1c (low level human venous whole blood K2 EDTA sample) and approximately 8.0% HbA1c (high level human venous whole blood K2 EDTA sample) using one lot of reagent on one DxC700 AU analyzer. Low and high pools were prepared by interdiluting human whole blood samples.
The whole blood samples were tested at a minimum of 5 levels for each interferent (Conjugated Bilirubin, Unconjugated Bilirubin, Lipemia, Ascorbic Acid, RF, Total Protein and Glucose) with 10 replicates tested per level on the DxC700 AU Analyzer to determine the magnitude of the interference.
No Significant Interference is recovery within 7% of the initial value (sample containing no interferent). Results in Table 1 show no significant interference up to the stated concentrations.
{23}------------------------------------------------
| Potential InterferingSubstance | Interference pool details | Interferent Level |
|---|---|---|
| Conjugated Bilirubin | Whole Blood Pools spiked withConjugated Bilirubin Stock Solution | 60 mg/dL |
| Unconjugated Bilirubin | Whole Blood Pools spiked withunconjugated Bilirubin StockSolution | 60 mg/dL |
| Lipemia | Whole Blood Pools spiked with20% w/v Intralipid | 500 mg/dL |
| Ascorbic Acid | Whole Blood Pools spiked withAscorbate | 300 mg/dL |
| RF | Whole Blood Pools spiked with RFpositive plasma | 1000 IU/ml |
| Total Protein | Whole Blood Pools spiked withHuman Serum Albumin | 21 g/dL |
| Glucose | Whole Blood Pools spiked withAnhydrous Glucose | 2000 mg/dL |
| Potential InterferingSubstance | Interference pool details | Interferent Level |
| Glyburide | Whole Blood Pools spikedwith Glyburide StockSolution | 0.12 mg/dL |
| Salicylic Acid | Whole Blood Pools spikedwith Salicylic Acid StockSolution | 4.76 mg/dL |
| Sitagliptin | Whole Blood Pools spikedwith Sitagliptin Phosphatestock solution | 0.2 mg/dL |
| Rosiglitazone | Whole Blood Pools spikedwith Rosiglitazone maleatestock solution | 0.33 mg/dL |
| Metformin | Whole Blood Pools spikedwith MetforminHydrochloride stocksolution | 5 mg/dL |
| Cyclosporine | Whole Blood Pools spikedwith Cyclosporine stocksolution | 0.5 mg/dL |
| Heparin | Whole Blood Pools spikedwith Sodium Heparin stocksolution | 5500 IU/L |
| Calcium Dobesilate | Whole Blood Pools spikedwith Calcium Dobsilatestock solution | 20 mg/dL |
| Metronidazole | Whole Blood Pools spikedwith Metronidazole stocksolution | 200 mg/dL |
| Levodopa | Whole Blood Pools spikedwith Levodopa Stocksolution | 20 mg/dL |
| Acetylsalicyclic acid | Whole Blood spiked withAcetylsalicyclic acid StockSolution | 1000 mg/dL |
| Acarbose | Whole blood spiked withAscarbose Stock Solution | 0.05 mg/dL |
| Acetaminophen | Whole blood spiked withAcetaminophen StockSolution | 26 mg/dL |
| Acetylcystein | Whole blood spiked withAcetylcystein StockSolution | 166 mg/dL |
| Ampicillin-Na | Whole blood spiked withAmpicillin-Na StockSolution | 1000 mg/dL |
| Cefoxitin | Whole blood spiked withCefoxin Stock Solution | 2500 mg/dL |
| Doxycyclin | Whole blood spiked withDoxycyclin Stock Solution | 50 mg/dL |
| Ibuprofen | Whole blood spiked withIbuprofen Stock Solution | 50 mg/dL |
| Methyldopa | Whole blood spiked withMethyldopa Stock Solution | 20 mg/dL |
| Phenylbutazone | Whole blood spiked withPhenylbutazone StockSolution | 53.5 mg/dL |
| Rifampicin | Whole blood spiked withRifampicin Stock Solution | 8 mg/dL |
| Theophylline | Whole blood spiked withTheophylline StockSolution | 10 mg/dL |
Table 17: Specifications and Summary of Endogenous Interference data
Drug Interference
Potential drug interfering substances were analyzed to determine their effect on HbA1c Advanced reagent . All drug interferences were tested at two % HbA1c concentrations, approximately 6.5% HbA1c (low level human venous whole blood EDTA sample) and approximately 8.0% HbA1c (high level human venous whole blood EDTA sample). Testing was completed using one lot of reagent on DxC 700 AU Analyzer.
Low and high pools were prepared by inter-diluting human whole blood samples. The two whole blood sample pools were spiked with each potential interfering drug to create test samples. Ten replicates of each test and reference sample containing no potential interferent) were tested on the DxC 700 AU to determine the magnitude of the interference. Each drug was tested to at least 5 times the highest drug concentration under therapeutic treatment.
No Significant Interference is recovery within 7% of the initial value (sample containing no interferent). Results outlined below showed no significant interference up to the stated concentrations.
{24}------------------------------------------------
Table 18: Specifications and Summary of Drug Interference data
{25}------------------------------------------------
Hemoglobin Derivative and Cross Reactants
Hemoglobin derivatives and potential interfering cross reactants were tested at two % HbA1c concentrations; approximately 6.5% HbA1c (low level human venous whole blood K2 EDTA sample) and approximately 8.0% HbA1c (high level human venous whole blood K2 EDTA sample) using one lot of reagent on one DxC700 AU analyzer. Low and high pools were prepared by interdiluting human whole blood samples.
Hemoglobin derivatives and potentially interfering cross reactants were tested at a minimum of 5 levels for: Labile Hemoglobin, Acetylated Hemoglobin, Carbamylated Hemoglobin, Glycated Albumin, HbA0 and HbA1a+b with 10 replicates tested per level on the DxC 700 AU analyzer to determine the magnitude of any interference effect.
No Significant Interference is recovery within 7% of the initial value (sample containing no interferent). Results in Table 19 showed no significant interference up to the stated concentrations.
{26}------------------------------------------------
Table 19: Specification and Summary of Hemoglobin Derivative and Cross Reactant Interferences
| Potential InterferingSubstance | Interference Pool details | Interferent Level |
|---|---|---|
| Labile Hemoglobin | Whole Blood Pools spiked withGlucose Stock Solution | 2000mg/dL |
| AcetylatedHemoglobin | Whole Blood Pools spiked withAcetylsalicylic acid Stock Solution | 0.5 mg/mL |
| CarbamylatedHemoglobin | Whole Blood Pools spiked withPotassium Cyanate StockSolution | 1.5 mg/mL |
| Glycated Albumin | Whole Blood Pools spiked withHuman Glycated Albumin StockSolution | 5mg/mL |
| HbA0 | Whole Blood Pools spiked withHbA0 Stock Solution | 12 mg/mL |
| HbA1a + 1b | Whole Blood Pools spiked withHbA1a +1b Stock Solution | 0.16 mg/mL |
Hemoglobin Variants
A hemoglobin variant study was performed to determine the presence of significant hemoglobin variant interference with major hemoglobin variants in the HbA1c Advanced assay. Results from singlicate measurements of samples were compared to results from a reference method, demonstrated to be free from hemoglobin interference; Trinity Biotech Hb9210 and Ultra2, Menarini HA8181V and TOSOH G8.
A minimum of 20 samples were tested for each variant (HbC, HbD, HbE, HbF, HbS and HbA2) on the DxC 700 AU Analyzer.
The following tables outline the number of samples of each variant, the range of % variant concentrations, %HbA1c concentrations and the relative % bias at 6.5% and 9.0% HbA1c.
{27}------------------------------------------------
Table 20: Hemoglobin Variant Sample Ranges
| HemoglobinVariant | Number ofSamples | Range in %VariantConcentration | Range in %HbA1cConcentration |
|---|---|---|---|
| HbC | 28 | 28.5 - 38.2 | 4.8 - 14.7 |
| HbD | 23 | 31.1 - 42.0 | 5.0 - 10.6 |
| HbE | 24 | 20.1 - 36.1 | 5.4 - 10.8 |
| HbF | 22 | 3.2 - 34.0 | 4.9 - 12.8 |
| HbS | 29 | 31.0 - 42.0 | 5.0 - 12.7 |
| HbA2 | 28 | 3.3 - 6.2 | 5.4 - 7.5 |
Table 21: Relative Bias Summary
| Hb Variant | Relative % bias (Range of % bias) observed relative toReference method | |
|---|---|---|
| HbA1c~6.5% HbA1c | HbA1c~9.0% HbA1c | |
| HbC | -2.57 (-4.30% to -1.80%) | -3.19 (-6.48% to 0.41%) |
| HbD | -0.77 (-4.81% to 2.99%) | -1.22 (-6.30% to -0.22%) |
| HbE | -1.12 (-9.16% to 2.48%) | 0.47 (-1.76% to 4.21%) |
| HbS | -1.18 (-2.17% to 3.04%) | -1.04 (-3.33% to 4.41%) |
| HbA2 | 0.48 (-1.92% to 5.60%) | 2.49 (-0.98% to 3.60%) |
| HbF | Specimens containing high amounts of HbF (> 7%) may yield lower than expected HbA1c Values |
{28}------------------------------------------------
Anticoaqulants
The data supports the use of the following blood collection tubes with the HbA1c Advanced reagent .
- K2 EDTA ●
- K3 EDTA ●
- Sodium Heparin ●
- Lithium Heparin .
Expected Values/Reference Interval
Beckman Coulter used evidence recommended by the American Diabetes Association Standards of (ADA), which may be used as an aid in the diagnosis of diabetes mellitus:
Table 1: HbA1c ranges recommended by the ADA as an aid in the diagnosis of diabetes mellitus 1,2,3
| HbA1c Advanced | ||
|---|---|---|
| NGSP % | IFCC mmol/mol | SuggestedDiagnosis |
| ≥ 6.5 | ≥48 | Diabetic |
| 5.7 – 6.4 | 39 - 47 | Pre-Diabetic |
| < 5.7 | <39 | Non-Diabetic |
Conclusion
The information and data in this 510(k) document demonstrate that the HbA1c Advanced reagent on the DxC700 AU Analyzer 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 HbA1c Advanced Reagent is substantially equivalent to the Biorad D-100™ HbA1c and D-100™HbA1c Calibrator Pack. It is therefore safe and effective for its intended use. The submitted information in this premarket notification is complete and supports a substantial equivalence decision.
§ 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.