HbA1c Advanced
Device Facts
| Record ID | K182651 |
|---|---|
| Device Name | HbA1c Advanced |
| Applicant | Beckman Coulter Ireland, Inc. |
| Product Code | PDJ · Clinical Chemistry |
| Decision Date | Jan 16, 2019 |
| Decision | SESE |
| Submission Type | Traditional |
| Regulation | 21 CFR 862.1373 |
| Device Class | Class 2 |
Indications for Use
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.
Device Story
HbA1c Advanced assay is an in vitro diagnostic reagent kit for use on the Beckman Coulter DxC700 AU Clinical Chemistry Analyzer. It processes human venous whole blood samples (K2-EDTA, K3-EDTA, Li-Heparin, Na-Heparin). The device uses an automated hemolysis step with tetradecyltrimethylammonium bromide (TTAB) to eliminate leukocyte interference. Total hemoglobin is measured colorimetrically (570/660 nm). HbA1c is measured via a turbidimetric immunoinhibition method (340/700 nm) where HbA1c antibodies form soluble complexes with sample HbA1c, and polyhaptens bind excess antibodies for turbidimetric measurement. The system calculates the HbA1c/Total Hemoglobin ratio, outputting results in mmol/mol (IFCC) or %HbA1c (DCCT/NGSP). Used in clinical laboratories by trained personnel; results assist clinicians in diagnosing diabetes and monitoring long-term glycemic control. The device provides standardized, precise quantification of HbA1c, aiding in patient management and risk assessment.
Clinical Evidence
Bench testing only. Precision evaluated per CLSI EP05-A3 over 20 days (n=2) across 3 lots and 3 instruments; total precision CVs ranged from 1.14% to 2.09% (NGSP). Linearity confirmed for 4-15% HbA1c (NGSP) and 20-140 mmol/mol (IFCC). Method comparison (n=138) against NGSP Secondary Reference Laboratory (HPLC) showed strong correlation (R=0.998, slope 0.990). Interference studies (CLSI EP07) confirmed no significant interference (recovery within 7%) for endogenous substances, drugs, hemoglobin derivatives, and common variants (HbC, D, E, S, A2). HbF >7% may cause lower results.
Technological Characteristics
Turbidimetric inhibition immunoassay. Reagents: Anti-human HbA1c antibody (sheep), HbA1c polyhapten, TTAB hemolyzing reagent. Analyzed on DxC 700 AU Clinical Chemistry Analyzer. Measures absorbance at 340/700 nm (HbA1c) and 570/660 nm (T-Hb). Standardized to IFCC reference material; NGSP certified. Liquid, ready-to-use format. Automated hemolysis.
Indications for Use
Indicated for quantitative determination of HbA1c in human venous whole blood to aid in diabetes mellitus diagnosis, monitor long-term glucose control in diabetic patients, and identify individuals at risk for developing diabetes.
Regulatory Classification
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.
Special Controls
(b) Classification. Class II (special controls). Hemoglobin A1c test systems must comply with the following special controls: 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%, 6.5%, 8.0% and 12% hemoglobin A1c. This testing must evaluate precision over a minimum of 20 days using at least 3 lots of the device and 3 instruments, as applicable. ii) Performance testing of device accuracy must include a minimum of 120 blood samples that span the measuring interval of the new device and compare results of the new device to results of the 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%. 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.
*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.
Predicate Devices
- D-100™ HbA1c, D-100™HbA1c Calibrator Pack (K151321)
Reference Devices
- Trinity Premier Hb9210
- Trinity Ultra Primus 2
- Menarini HA8180V (k162822)
- Tosoh G8
Related Devices
- K120199 — GLYCOSYLATED HEMOGLOBIN ASSAY; CLINICAL CHEMISTRY · Beckman Coulter, Inc. · Oct 12, 2012
- K173909 — Dimension Hemoglobin A1c Assay · Siemens Healthcare Diagnostics, Inc. · Jul 13, 2018
- K121492 — UNICEL DXC SYNCHRON SYSTEMS HEMOGLOBIN ALC-(HBA1C-) REAGENT · Beckman Coulter, Inc. · Sep 11, 2012
Submission Summary (Full Text)
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# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY ASSAY ONLY TEMPLATE
A. 510(k) Number:
k182651
B. Purpose for Submission:
New Device
C. Measurand:
Glycosylated Hemoglobin (HbA1c)
D. Type of Test:
Quantitative turbidimetric inhibition immunoassay
E. Applicant:
Beckman Coulter Ireland Inc.
F. Proprietary and Established Names:
HbA1c Advanced
G. Regulatory Information:
| Regulation Description | Product Code | Device Class | Regulation | Panel |
| --- | --- | --- | --- | --- |
| Hemoglobin A1c Test System | PDJ | II | 21 CFR § 862.1373 | Chemistry, 75 |
| Glycosylated Hemoglobin Assay | LCP | II | 21 CFR 864.7470 | Hematology, 81 |
H. Intended Use:
1. Intended use(s):
See Indications for use below.
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2. Indication(s) for use:
The HbA1c (Hemoglobin A1c) 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.
3. Special conditions for use statement(s):
For prescription use only.
For in vitro diagnostic use.
This assay is designed only for the measurement of mmol/mol HbA1c (IFCC) and %HbA1c (NGSP). Individual results for T-Hb and A1c concentration should not be reported.
This test should not replace glucose testing for patients with Type 1 diabetes, pediatric patients or pregnant women.
In cases of rapidly evolving Type 1 diabetes, the increase of HbA1c values might be delayed compared to the acute increase in glucose concentrations. In these conditions, diabetes mellitus must be diagnosed based on plasma glucose concentration and/or the typical clinical symptoms.
HbA1c should not be used to diagnose diabetes during pregnancy or to diagnose gestational diabetes. HbA1c reflects the average blood glucose levels over the preceding 3 months (the average life of a red blood cell), and therefore may be falsely low during pregnancy or any other condition associated with recent onset of hyperglycemia.
The HbA1c Advanced assay allows its use in patients with hemoglobin variants S, C, D and E traits. However, hemoglobinopathies might affect the red blood cell turnover or the in vivo glycation rates, and these patients may suffer from anemia and undergo transfusion. Therefore, results reported correctly by the assay may not reflect the same level of glycemic control as would be expected in patients with normal hemoglobin. In these cases, even analytically correct results do not reflect the same level of glycemic control that would be expected in patients with normal hemoglobin. Whenever, it is suspected that the presence of a Hb variant (e.g. HbSS, HbCC or HbSC) affects the correlation between the HbA1c value and glycemic control, HbA1c must not be used for the diagnosis of diabetes mellitus.
This test is not intended for analysis of samples collected from newborns.
Glycated HbF is not detected by the assay as it does not contain the β-chain that
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characterizes HbA1c. However, HbF is measured in the T-Hb assay and as a consequence, specimens containing high amounts of HbF may result in lower than expected mmol/mol HbA1c values (IFCC) and% HbA1c values (DCCT/NGSP). HbA1c results are invalid for patients with abnormal amounts of HbF including those with known Hereditary Persistence of Fetal Hemoglobin. The following warning is included in the labeling;
"WARNING: The Hemoglobin A1c assay has significant negative interference from Hemoglobin F (HbF). HbA1c results are invalid for patients with abnormal amounts of HbF, including those with known Hereditary Persistence of Fetal Hemoglobin. Refer to Limitations section."
HbA1c should not be used to diagnose or monitor diabetes in patients with any conditions resulting in decreased erythrocyte survival, decrease in mean erythrocyte age or abnormal red blood cell turnover (e.g., hemolytic anemia, including hereditary spherocytosis, iron deficiency, thalassemias, malignancies, recovery from acute blood loss and severe chronic hepatic and renal disease). Shortened red cell survival time will reduce the exposure of red cells to glucose, with a resultant decrease in HbA1c values. The altered red blood cell turnover interferes with the relationship between mean blood glucose and HbA1c values.
4. Special instrument requirements:
Beckman Coulter DxC 700 AU Clinical Chemistry Analyzer (k161837)
I. Device Description:
The HbA1c Advanced reagent kit is provided in a liquid format and is ready to use. It contains HbA1c R1 and HbA1c R2, Total Hemoglobin R1 and Hemolyzing reagent R1.
| HbA1c R1 | Anti-human HbA1c Antibody (sheep) |
| --- | --- |
| | MES (2-morpholino-ethanesulphonic acid) Buffer |
| | TRIS (tris(hydroxymethyl)aminomethane) Buffer (pH 6.2) |
| HbA1c R2 | HbA1c Polyhapten |
| | MES (2-morpholino-ethanesulphonic acid) Buffer |
| | TRIS (tris(hydroxymethyl)aminomethane) Buffer (pH 6.2) |
| Total Hemoglobin R1 | Phosphate Buffer, pH 7.4 |
| Hemolyzing reagent R1 | 0.9% Tetradecyltrimethylammonium bromide |
The HbA1c calibrator is supplied with the reagent, in a liquid, ready to use format and contains 5 x 2 mL calibrator levels (levels 2 - 6). The sample hemolysis is automated on the DxC 700 AU Clinical Chemistry analyzer.
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J. Substantial Equivalence Information:
1. Predicate device name(s):
Bio-Rad D-100™ HbA1c
2. Predicate 510(k) number(s):
k151321
3. Comparison with predicate:
| Similarities | | |
| --- | --- | --- |
| Item | Candidate Device HbA1c Advanced | Predicate Device Bio-Rad D-100™ HbA1c k151321 |
| Intended Use | 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. | Same |
| Specimen Type | Venous whole blood
K2-EDTA
K3-EDTA
Lithium Heparin
Sodium Heparin | Same |
| Reporting Units | NGSP Equivalent Units (% HbA1c)
IFCC Equivalent Units (mmol/mol) | Same |
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| Differences | | |
| --- | --- | --- |
| Item | Candidate Device HbA1c Advanced | Predicate Device Bio-Rad D-100™ HbA1c k151321 |
| Platform | DxC 700 AU Clinical Chemistry Analyzer | D-100TM Hemoglobin Testing System |
| Methodology | Quantitative turbidimetric inhibition immunoassay | Ion-exchange Quantitative high performance liquid chromatography (HPLC) |
| Measuring Range | HbA1c: 4.0 – 15% HbA1c (NGSP) 20 – 140 mmol/mol (IFCC) | HbA1c: 3.5 to 20% HbA1c (NGSP) 15 - 195 mmol/mol (IFCC) |
# K. Standard/Guidance Document Referenced (if applicable):
CLSI EP05-A3: Evaluation of Precision of Quantitative Measurement Procedures; Approved Guideline - Third Edition
CLSI EP06-A: Evaluation of the Linearity of Quantitative Measurement Procedures: A Statistical Approach; Approved Guideline
CLSI EP07-A2: Interference Testing in Clinical Chemistry; Approved Guideline - Second Edition
CLSI EP-37 1st Edition: Supplemental tables for Interference testing in Clinical Chemistry - First edition
CLSI EP25-A: Evaluation of Stability of In Vitro Diagnostic Reagents; Approved Guideline
# L. Test Principle:
The HbA1c Advanced assay is a turbidimetric inhibition immunoassay. The DxC 700 AU Clinical Chemistry analyzer automatically performs the whole blood hemolysis.
Tetradecyltrimethylammonium bromide (TTAB) in the hemolyzing reagent eliminates interference from leukocytes. The hemolyzed venous whole blood is then added to both the Total Hemoglobin and HbA1c assay cuvettes according to their defined assay parameters.
Total Hemoglobin reagent (R1) is used to measure Total Hemoglobin (T-Hb) concentration by a colorimetric method. Change in absorbance is measured at $570 / 660\mathrm{nm}$ .
HbA1c reagent (R1 and R2) is used to measure hemoglobin A1c (A1c) concentration by a turbidimetric immunoinhibition method. In the reaction, hemoglobin A1c antibodies in the reagent 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
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agglutinated complex is measured turbidimetrically. Change in absorbance is measured at 340/700 nm.
The concentrations of both T-Hb and A1c are determined, and are used in the calculation of the reported HbA1c (A1c/T-Hb ratio), which is expressed either as mmol/mol (IFCC) or % (DCCT/NGSP).
## M. Performance Characteristics (if/when applicable):
### 1. Analytical performance:
#### a. Precision/Reproducibility:
Precision of the HbA1c Advanced reagent on the DxC 700 AU Clinical Chemistry Analyzer was evaluated based on CLSI guideline EP05-A3, Evaluation of Precision Performance of Quantitative Measurement Methods; Approved Guideline - Third Edition. Studies were carried out at a single site by three operators over 20 days on three DxC 700 AU systems using three lots of HbA1c Advanced reagent, four levels of HbA1c K2 EDTA human venous whole blood patient samples at HbA1c concentrations of approximately 5.0%, 6.5%, 8.0% and 12%, one level of K2 EDTA human venous whole blood spiked with human based high HbA1c material at a HbA1c concentration of approximately 14% and two levels of control materials (QC) resulting in 720 measurements for each sample. The summary of precision performance results are shown in the tables below.
DxC 700 AU Clinical Chemistry Analyzer Instrument 1, % HbA1c (NGSP) units
| Sample ID | Mean HbA1c % | Repeatability | | Between Run | | Between Day | | Between Lot | | Total Precision | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | | SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% |
| Pool 1 | 5.06 | 0.04 | 0.88 | 0.04 | 0.80 | 0.03 | 0.64 | 0.05 | 0.92 | 0.08 | 1.63 |
| Pool 2 | 6.72 | 0.07 | 1.01 | 0.04 | 0.60 | 0.04 | 0.65 | 0.06 | 0.93 | 0.11 | 1.64 |
| Pool 3 | 8.06 | 0.06 | 0.77 | 0.04 | 0.47 | 0.06 | 0.73 | 0.09 | 1.06 | 0.13 | 1.57 |
| Pool 4 | 11.70 | 0.09 | 0.79 | 0.06 | 0.48 | 0.10 | 0.84 | 0.02 | 0.19 | 0.15 | 1.26 |
| Pool 5 | 14.02 | 0.10 | 0.74 | 0.07 | 0.49 | 0.10 | 0.72 | 0.04 | 0.32 | 0.17 | 1.19 |
| QC 1 | 5.32 | 0.06 | 1.19 | 0.04 | 0.84 | 0.03 | 0.66 | 0.07 | 1.34 | 0.11 | 2.08 |
| QC 2 | 9.88 | 0.08 | 0.77 | 0.05 | 0.48 | 0.06 | 0.65 | 0.11 | 1.07 | 0.15 | 1.54 |
DxC 700 AU Clinical Chemistry Analyzer Instrument 2, % HbA1c (NGSP) units
| Sample ID | Mean HbA1c % | Repeatability | | Between Run | | Between Day | | Between Lot | | Total Precision | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | | SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% |
| Pool 1 | 5.03 | 0.06 | 1.15 | 0.02 | 0.37 | 0.03 | 0.57 | 0.05 | 1.02 | 0.08 | 1.68 |
| Pool 2 | 6.69 | 0.08 | 1.14 | 0.01 | 0.20 | 0.03 | 0.50 | 0.06 | 0.83 | 0.10 | 1.51 |
| Pool 3 | 8.05 | 0.07 | 0.86 | 0.04 | 0.48 | 0.04 | 0.49 | 0.06 | 0.70 | 0.11 | 1.31 |
| Pool 4 | 11.69 | 0.08 | 0.71 | 0.04 | 0.33 | 0.07 | 0.61 | 0.07 | 0.56 | 0.13 | 1.14 |
| Pool 5 | 14.04 | 0.11 | 0.75 | 0.08 | 0.58 | 0.05 | 0.34 | 0.10 | 0.69 | 0.17 | 1.22 |
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| Sample ID | Mean HbA1c % | Repeatability | | Between Run | | Between Day | | Between Lot | | Total Precision | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | | SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% |
| QC 1 | 5.28 | 0.06 | 1.23 | 0.02 | 0.41 | 0.04 | 0.75 | 0.07 | 1.32 | 0.11 | 1.99 |
| QC 2 | 9.88 | 0.08 | 0.77 | 0.04 | 0.42 | 0.05 | 0.54 | 0.09 | 0.95 | 0.14 | 1.40 |
DxC 700 AU Clinical Chemistry Analyzer Instrument 3, % HbA1c (NGSP) units
| Sample ID | Mean HbA1c % | Repeatability | | Between Run | | Between Day | | Between Lot | | Total Precision | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | | SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% |
| Pool 1 | 5.03 | 0.04 | 0.83 | 0.02 | 0.45 | 0.04 | 0.70 | 0.05 | 1.02 | 0.08 | 1.55 |
| Pool 2 | 6.66 | 0.06 | 0.82 | 0.02 | 0.31 | 0.03 | 0.48 | 0.09 | 1.31 | 0.11 | 1.65 |
| Pool 3 | 7.98 | 0.06 | 0.73 | 0.05 | 0.58 | 0.04 | 0.47 | 0.14 | 1.68 | 0.16 | 1.98 |
| Pool 4 | 11.68 | 0.08 | 0.71 | 0.03 | 0.21 | 0.08 | 0.67 | 0.06 | 0.54 | 0.13 | 1.14 |
| Pool 5 | 13.99 | 0.09 | 0.66 | 0.06 | 0.44 | 0.08 | 0.56 | 0.09 | 0.66 | 0.17 | 1.18 |
| QC 1 | 5.25 | 0.05 | 0.93 | 0.05 | 0.97 | 0.03 | 0.60 | 0.07 | 1.37 | 0.11 | 2.01 |
| QC 2 | 9.72 | 0.06 | 0.65 | 0.03 | 0.36 | 0.06 | 0.65 | 0.14 | 1.45 | 0.17 | 1.75 |
DxC 700 AU Clinical Chemistry Analyzer Instruments Combined, % HbA1c (NGSP) units
| Sample ID | Mean HbA1c % | Repeat- ability | | Between Run | | Between Day | | Between Lot | | Between Instrument | | Total Precision | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | | SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% |
| Pool 1 | 5.07 | 0.05 | 0.96 | 0.03 | 0.57 | 0.03 | 0.64 | 0.05 | 0.99 | 0.04 | 0.74 | 0.09 | 1.78 |
| Pool 2 | 6.72 | 0.07 | 1.00 | 0.03 | 0.41 | 0.04 | 0.55 | 0.07 | 1.05 | 0.04 | 0.56 | 0.11 | 1.69 |
| Pool 3 | 8.07 | 0.06 | 0.79 | 0.04 | 0.51 | 0.05 | 0.58 | 0.10 | 1.22 | 0.05 | 0.57 | 0.14 | 1.74 |
| Pool 4 | 11.71 | 0.09 | 0.74 | 0.04 | 0.36 | 0.08 | 0.71 | 0.05 | 0.46 | 0.04 | 0.32 | 0.14 | 1.22 |
| Pool 5 | 14.03 | 0.10 | 0.72 | 0.07 | 0.51 | 0.08 | 0.56 | 0.08 | 0.58 | 0.02 | 0.12 | 0.17 | 1.20 |
| QC 1 | 5.29 | 0.06 | 1.12 | 0.04 | 0.78 | 0.04 | 0.67 | 0.07 | 1.34 | 0.03 | 0.50 | 0.11 | 2.09 |
| QC 2 | 9.84 | 0.07 | 0.73 | 0.04 | 0.43 | 0.06 | 0.61 | 0.12 | 1.17 | 0.07 | 0.70 | 0.17 | 1.72 |
DxC 700 AU Clinical Chemistry Analyzer Instrument 1, mmol/mol (IFCC) units
| Sample ID | Mean HbA1c % | Repeatability | | Between Run | | Between Day | | Between Lot | | Total Precision | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | | SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% |
| Pool 1 | 31.84 | 0.48 | 1.51 | 0.46 | 1.43 | 0.34 | 1.08 | 0.51 | 1.60 | 0.90 | 2.84 |
| Pool 2 | 49.92 | 0.75 | 1.49 | 0.45 | 0.90 | 0.48 | 0.97 | 0.68 | 1.36 | 1.21 | 2.41 |
| Pool 3 | 64.65 | 0.68 | 1.06 | 0.40 | 0.61 | 0.65 | 1.01 | 0.93 | 1.43 | 1.38 | 2.14 |
| Pool 4 | 104.38 | 1.37 | 1.31 | 0.00 | 0.00 | 1.07 | 1.03 | 0.24 | 0.23 | 1.76 | 1.68 |
| Pool 5 | 129.76 | 1.13 | 0.87 | 0.74 | 0.57 | 1.11 | 0.86 | 0.49 | 0.37 | 1.82 | 1.40 |
| QC 1 | 34.65 | 0.70 | 2.01 | 0.44 | 1.28 | 0.41 | 1.18 | 0.77 | 2.22 | 1.20 | 3.46 |
| QC 2 | 84.53 | 0.83 | 0.98 | 0.51 | 0.61 | 0.69 | 0.82 | 1.15 | 1.36 | 1.66 | 1.96 |
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DxC 700 AU Clinical Chemistry Analyzer Instrument 2, mmol/mol (IFCC) units
| Sample ID | Mean HbA1c % | Repeatability | | Between Run | | Between Day | | Between Lot | | Total Precision | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | | SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% |
| Pool 1 | 31.51 | 0.63 | 2.01 | 0.37 | 1.19 | 0.24 | 0.77 | 0.57 | 1.81 | 0.96 | 3.05 |
| Pool 2 | 49.59 | 0.81 | 1.63 | 0.21 | 0.42 | 0.37 | 0.74 | 0.59 | 1.18 | 1.08 | 2.18 |
| Pool 3 | 64.48 | 0.76 | 1.18 | 0.43 | 0.66 | 0.43 | 0.67 | 0.62 | 0.96 | 1.15 | 1.79 |
| Pool 4 | 104.29 | 0.91 | 0.88 | 0.40 | 0.38 | 0.78 | 0.75 | 0.73 | 0.70 | 1.46 | 1.40 |
| Pool 5 | 129.92 | 1.15 | 0.88 | 0.85 | 0.66 | 0.56 | 0.43 | 1.06 | 0.82 | 1.87 | 1.44 |
| QC 1 | 34.24 | 0.72 | 2.09 | 0.25 | 0.74 | 0.42 | 1.23 | 0.76 | 2.23 | 1.16 | 3.38 |
| QC 2 | 84.52 | 0.83 | 0.98 | 0.46 | 0.54 | 0.58 | 0.69 | 1.03 | 1.22 | 1.52 | 1.80 |
DxC 700 AU Clinical Chemistry Analyzer Instrument 3, mmol/mol (IFCC) units
| Sample ID | Mean HbA1c % | Repeatability | | Between Run | | Between Day | | Between Lot | | Total Precision | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | | SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% |
| Pool 1 | 31.46 | 0.46 | 1.41 | 0.38 | 1.18 | 0.42 | 1.31 | 0.58 | 1.78 | 0.93 | 2.88 |
| Pool 2 | 49.27 | 0.61 | 1.21 | 0.23 | 0.46 | 0.35 | 0.70 | 0.97 | 1.93 | 1.22 | 2.42 |
| Pool 3 | 63.68 | 0.65 | 1.00 | 0.52 | 0.79 | 0.41 | 0.63 | 1.49 | 2.29 | 1.75 | 2.69 |
| Pool 4 | 104.18 | 0.92 | 0.87 | 0.26 | 0.25 | 0.85 | 0.81 | 0.70 | 0.66 | 1.46 | 1.39 |
| Pool 5 | 129.37 | 1.02 | 0.79 | 0.67 | 0.52 | 0.86 | 0.66 | 1.01 | 0.87 | 1.81 | 1.39 |
| QC 1 | 33.83 | 0.54 | 1.58 | 0.55 | 1.61 | 0.35 | 1.02 | 0.79 | 2.32 | 1.16 | 3.40 |
| QC 2 | 82.77 | 0.69 | 0.83 | 0.38 | 0.46 | 0.69 | 0.83 | 1.55 | 1.86 | 1.87 | 2.24 |
DxC 700 AU Clinical Chemistry Analyzer Instruments Combined, mmol/mol (IFCC) units
| Sample ID | Mean HbA1c % | Repeat- ability | | Between Run | | Between Day | | Between Lot | | Between Instrument | | Total Precision | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | | SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% |
| Pool 1 | 31.91 | 0.53 | 1.66 | 0.41 | 1.27 | 0.34 | 1.08 | 0.55 | 1.73 | 0.42 | 1.31 | 1.02 | 3.20 |
| Pool 2 | 49.95 | 0.72 | 1.45 | 0.31 | 0.63 | 0.41 | 0.81 | 0.76 | 1.53 | 0.41 | 0.82 | 1.24 | 2.49 |
| Pool 3 | 64.75 | 0.70 | 1.08 | 0.45 | 0.69 | 0.51 | 0.79 | 1.07 | 1.66 | 0.50 | 0.77 | 1.53 | 2.37 |
| Pool 4 | 104.51 | 1.09 | 1.04 | 0.24 | 0.23 | 0.91 | 0.87 | 0.60 | 0.57 | 0.42 | 0.40 | 1.61 | 1.54 |
| Pool 5 | 129.84 | 1.10 | 0.85 | 0.76 | 0.58 | 0.87 | 0.67 | 0.89 | 0.69 | 0.19 | 0.14 | 1.84 | 1.42 |
| QC 1 | 34.32 | 0.66 | 1.91 | 0.43 | 1.26 | 0.39 | 1.15 | 0.78 | 2.26 | 0.30 | 0.87 | 1.21 | 3.52 |
| QC 2 | 84.10 | 0.78 | 0.93 | 0.46 | 0.54 | 0.66 | 0.78 | 1.26 | 1.50 | 0.76 | 0.90 | 1.85 | 2.20 |
b. Linearity/assay reportable range:
Linearity testing was conducted based on CLSI EP06-A, Evaluation of the Linearity of Quantitative Measurement Procedures: A Statistical Approach; Approved Guideline. A dilution series consisting of ten levels across the assay range was prepared by mixing high HbA1c and low HbA1c venous whole blood pools. Samples levels evaluated were 3.94, 4.67, 6.14, 7.60, 9.07, 10.54, 12.00, 13.47, 14.94 and 15.37% HbA1c (19.51, 29.53, 43.56, 59.59, 75.62, 91.65, 107.68, 123.71, 139.74 and 144.49 mmol/mol HbA1c) and each was run in quadruplicate with two reagent lots on
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the DxC 700 AU Clinical Chemistry Analyzer. The measured values were compared to the expected values. The regression parameters (slope, intercept, and r) are shown below for one representative lot:
NGSP:
| Slope | Intercept | R | Concentration Range Tested |
| --- | --- | --- | --- |
| 1.030 | -0.14 | 1.000 | 3.94 to 15.37 % HbA1c |
IFCC:
| Slope | Intercept | R | Concentration Range Tested |
| --- | --- | --- | --- |
| 1.020 | -0.64 | 1.000 | 19.51 to 144.49 mmol/mol HbA1c |
The results of the linearity study support the claimed assay measuring range of 4-15% HbA1c (20 - 140 mmol/mol HbA1c (IFCC)).
c. Traceability, Stability, Expected values (controls, calibrators, or methods):
The HbA1c Advanced assay standardization is traceable to the International Federation of Clinical Chemistry (IFCC) reference material. The HbA1c Advanced assay is certified through the National Glycohemoglobin Standardization Program (NGSP). The NGSP certification expires in one year. See NGSP website for current certification at http://www.ngsp.org. The International Federation of Clinical Chemistry (IFCC) units of mmol/mol are calculated using the Master Equation:
$$
\mathrm{NGSP}(\%) = 0.09145 \times \mathrm{IFCC}(\mathrm{mmol/mol}) + 2.152
$$
$$
\mathrm{IFCC} \text{Formula} = \frac{A1c}{T - Hb} \times 1000 = HbA1c \text{ (mmol/mol)}
$$
Results for calculated HbA1c may be reported from the DxC 700 AU analyzer in either NGSP (%) units or IFCC (mmol/mol).
d. Detection limit:
The claimed measuring range of 4-15% HbA1c (NGSP) and 20 - 140 mmol/mol HbA1c (IFCC) for HbA1c Advanced reagent is based on the linearity studies above in M.1.b.
e. Analytical specificity:
Testing to determine the interference bias of various endogenous and exogenous interferents on the HbA1c Advanced Assay was performed according to CLSI EP07-A2, Interference Testing in Clinical Chemistry; Approved Guideline - Second Edition; CLSI EP37 Supplement Tables for Interference Testing in Clinical Chemistry, 1st Edition was used in conjunction with EP07-A2.
The interfering substances analyzed were tested at two % HbA1c concentrations; approximately 6.5% HbA1c (low level human venous whole blood K2 EDTA
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sample) and approximately 8.0% HbA1c (high level human venous whole blood K2 EDTA sample) using one lot of reagent on one DxC 700 AU Clinical Chemistry Analyzer.
The whole blood samples were tested at a minimum of five concentration levels for each interferent with 10 replicates tested per level on the DxC 700 AU Clinical Chemistry Analyzer to determine the magnitude of the interference. The difference in mean recovery of the samples with and without the potential interfering substances was determined.
The sponsor defines significant interference as ≥ ±7% change in the HbA1c measurements from the initial value (sample containing no interferent). The following substances demonstrated no significant interference at the concentrations described below:
Endogenous Interference Summary
| Potential Interfering Substance | Highest Concentration Tested without Significant Interference |
| --- | --- |
| Conjugated Bilirubin | 60 mg/dL |
| Unconjugated Bilirubin | 60 mg/dL |
| Lipemia (Intralipid) | 500 mg/dL |
| Ascorbic Acid | 300 mg/dL |
| Rheumatoid Factor | 1000 IU/mL |
| Total Protein | 21 g/dL |
| Glucose | 2000 mg/dL |
Exogenous Interference Summary
| Potential Interfering Substance | Highest Concentration Tested without Significant Interference |
| --- | --- |
| Glyburide | 0.12 mg/dL |
| Salicyclic Acid | 4.76 mg/dL |
| Sitagliptin | 0.2 mg/dL |
| Rosiglitazone | 0.33 mg/dL |
| Metformin | 5 mg/dL |
| Cyclosporine | 0.5 mg/dL |
| Heparin | 5500 IU/L |
| Calcium Dobesilate | 20 mg/dL |
| Metronidazole | 200 mg/dL |
| Levodopa | 20 mg/dL |
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| Potential Interfering Substance | Highest Concentration Tested without Significant Interference |
| --- | --- |
| Acetylsalicyclic acid | 1000 mg/dL |
| Acarbose | 0.05 mg/dL |
| Acetaminophen | 26 mg/dL |
| Acetylcystein | 166 mg/dL |
| Ampicillin-Na | 1000 mg/dL |
| Cefoxitin | 2500 mg/dL |
| Doxycyclin | 50 mg/dL |
| Ibuprofen | 50 mg/dL |
| Methylidopa | 20 mg/dL |
| Phenylbutazone | 53.5 mg/dL |
| Rifampicin | 8 mg/dL |
| Theophylline | 10 mg/dL |
## Cross reactivity with Hemoglobin derivatives
Hemoglobin derivatives and potential cross reactants were tested at two $\% \mathrm{HbA1c}$ concentrations, approximately $6.5\%$ and $8.0\%$ HbA1c, using one lot of reagent on one DxC 700 AU Clinical Chemistry Analyzer. Hemoglobin derivatives and potential cross reactants were tested at a minimum of five concentration levels with 10 replicates tested per level to determine the magnitude of any interference effect. The sponsor defined significant interference as recovery $>7\%$ of the initial value (sample containing no cross reactant).
The following substances demonstrated no significant cross reactivity at the concentrations described below:
| Potential Cross reacting Substance | Highest Concentration Level without Significant Cross Reactivity |
| --- | --- |
| Labile Hemoglobin | 2000 mg/dL (Glucose) |
| Acetylated Hemoglobin | 0.5 mg/mL (Acetylsalicylic acid) |
| Carbamylated Hemoglobin | 1.5 mg/mL (Potassium Cyanate) |
| Glycated Albumin | 5mg/mL |
| HbA0 | 12 mg/mL |
| HbA1a + b | 0.16 mg/mL |
## Hemoglobin Variant Interference
A study was performed to assess hemoglobin variant interference with major hemoglobin variants in the HbA1c Advanced assay. Results from singleate measurements of K2EDTA human venous whole blood samples tested using the HbA1c Advanced assay on the DxC 700 AU Clinical Chemistry Analyzer were compared to results from a FDA cleared method demonstrated to be free from
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hemoglobin interference; Trinity Premier Hb9210 and Trinity Ultra Primus 2 were used to verify %HbA1c concentrations in samples containing HbC, HbD, HbE, HbS, and HbA2 variants. The Menarini HA8180V and Tosoh G8 were used to verify %HbA1c concentrations in samples containing HbF variant. Significant interference was defined by the sponsor as $\geq \pm 7\%$ change in HbA1c value in the presence of the hemoglobin variant relative to control. The following tables show the samples that were measured and the range of biases that were observed.
Hemoglobin Variant Sample Ranges
| Hb Variant | Number of Samples | % Concentration of Variant in Sample | %HbA1c Concentration Range |
| --- | --- | --- | --- |
| 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 |
| HbS | 29 | 31.0 – 42.0 | 5.0 - 12.7 |
| HbA2 | 28 | 3.3 - 6.2 | 5.4 - 7.5 |
| HbF | 22 | 3.2 – 34.0 | 4.9 - 12.8 |
Relative Bias Summary
| Hb Variant | Relative % Bias (Range of % Bias) observed relative to Comparator Method | |
| --- | --- | --- |
| | HbA1c ~6.5% A1c | HbA1c ~9% A1c |
| 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 | |
The results show there is no significant interference for HbS ($\leq 42.0\%$), HbC ($\leq 38.2\%$), HbD ($\leq 42.0\%$), HbE ($\leq 36.1\%$), and HbA2 ($\leq 6.2\%$). The labeling states:
"WARNING: The Hemoglobin A1c assay has significant negative interference from Hemoglobin F (HbF). HbA1c results are invalid for patients with abnormal amounts of HbF, including those with known Hereditary Persistence of Fetal Hemoglobin. Refer to Limitations section."
f. Assay cut-off:
Not applicable.
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2. Comparison studies:
a. Method comparison with predicate device:
Method comparison testing was performed using 138 frozen human venous whole blood samples in K2EDTA with values spanning the assay range of 4.70 to 14.2% HbA1c. Samples were evaluated using the HbA1c Advanced assay on the DxC 700 AU Clinical Chemistry Analyzer. The HbA1c Advanced assay first replicate results were compared to results obtained for the same samples from testing at an NGSP secondary reference laboratory (SRL) on the Menarini HA8180V, a HPLC method (cleared in k162822). The distribution of samples spanning the measuring interval follows:
Distribution of Samples
| % HbA1c Level | n | % of Samples tested |
| --- | --- | --- |
| <5 | 6 | 4.35% |
| 5-6 | 18 | 13.04% |
| 6-6.5 | 32 | 23.19% |
| 6.5-7 | 35 | 25.36% |
| 7-8 | 25 | 18.12% |
| 8-9 | 12 | 8.7% |
| >9 | 10 | 7.25% |
| Total | 138 | 100% |
Deming (weighted) and Passing-Bablok regression analyses were performed for the HbA1c Advanced assay versus the comparator method. A summary of the results are provided below:
NGSP
| | Slope | 95 % CI | y-intercept | 95% CI | R |
| --- | --- | --- | --- | --- | --- |
| Deming | 0.990 | (0.978-1.002) | 0.01 | (-0.070-0.089) | 0.998 |
| Passing-Bablok | 0.980 | (0.964-0.992) | 0.090 | (-0.006-0.187) | 0.998 |
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Scatter Plot using Deming Fit, %HbA1c, HbA1c Advanced vs. NGSP SRL
Scatter Plot using Passing-Bablok Fit, %HbA1c, HbA1c Advanced vs. NGSP SRL
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Scatter Plot using Deming Fit, %HbA1c, HbA1c Advanced vs. IFCC SRL
Scatter Plot using Passing-Bablok Fit, %HbA1c, HbA1c Advanced vs. IFCC SRL
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Summary of Bias Estimation table (medical decision points) Weighted Deming Analysis
Bias Estimation:
| %HbA1c | Automated (Whole Blood) | |
| --- | --- | --- |
| | Bias | % Bias |
| 5.00 | -0.04 | -0.80 |
| 6.50 | 0.06 | -0.85 |
| 8.00 | -0.07 | -0.88 |
| 12.00 | -0.11 | -0.92 |
# Total Error calculations and estimation
The bias estimation values determined in the method comparison study and precision estimates determined in the precision study were used to determine the total error at each of the levels listed in the tables below. Total error was calculated by the following equation:
$\% \mathrm{TE} = |\% \mathrm{Bias}| + (1.96 \times \% \mathrm{CV}) \times (1 + \% \mathrm{Bias} / 100)$
Total Error Summary:
| %HbA1c | % Bias | %CV | %TE |
| --- | --- | --- | --- |
| 5.00 | -0.80 | 1.78 | 4.3 |
| 6.50 | -0.85 | 1.69 | 4.2 |
| 8.00 | -0.88 | 1.74 | 4.2 |
| 12.00 | -0.92 | 1.22 | 3.3 |
# b. Matrix comparison:
Testing was performed to validate the use of different anticoagulants in primary collection tubes for the HbA1c Advanced reagent on the DxC 700 AU Clinical Chemistry Analyzer. The study was carried out using one lot of HbA1c Advanced and K2 EDTA, K3 EDTA, Lithium Heparin and Sodium Heparin anticoagulant tube types. K2 EDTA values were used as the reference method for K3 EDTA, Lithium Heparin and Sodium Heparin.
Fifty native venous human whole blood specimens were drawn from each donor into each of the intended anticoagulant tube types. Values of K2-EDTA whole blood, ranging from $4.86\%$ HbA1c to $9.83\%$ HbA1c, were compared with values for K3-EDTA whole blood, lithium heparin whole blood and sodium heparin whole blood, yielding the following regression results:
| Anticoagulant | Deming Regression Analysis |
| --- | --- |
| K3-EDTA | y = 1.000x + 0.043 ; r = 0.997 |
| Lithium Heparin | y = 1.004x + 0.013; r = 0.994 |
| Sodium Heparin | y = 1.002x - 0.009; r = 0.997 |
These results support the use of the HbA1c Advanced assay with samples collected in
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K2-EDTA, K3-EDTA, Lithium Heparin or Sodium Heparin tubes.
3. Clinical studies:
a. Clinical Sensitivity:
Not applicable.
b. Clinical specificity:
Not applicable.
c. Other clinical supportive data (when a. and b. are not applicable):
Not applicable.
4. Clinical cut-off:
Not applicable.
5. Expected values/Reference range:
Hemoglobin A1c Expected Values:
| Hemoglobin A1c | | Suggested Diagnosis |
| --- | --- | --- |
| NGSP % | IFCC mmol/mol | |
| >6.5 | >48 | Diabetic |
| 5.7 – 6.4 | 39-47 | Pre-Diabetic |
| <5.7 | <39 | Non-Diabetic |
The expected HbA1c range for non-diabetic adults is 4-6%.
The following references were cited in the labeling:
- American Diabetes Association. Position Statement: Standards of medical care in diabetes – In: Diabetes Care 2018; 41 (Suppl 1): S13-S27.
- Panteghini M, John WG. Implementation of Hemoglobin A1c results traceable to the IFCC reference system: the way forward. Clin Chem Lab Med 2007; 45(8): 942-944.
N. Proposed Labeling:
The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10 and the special controls for this device type under 21 CFR 862.1373.
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O. Conclusion:
The submitted information in this premarket notification is complete and supports a substantial equivalence decision.
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