(294 days)
The cobas c 513 clinical chemistry analyzer is a fully automated, standalone clinical chemistry analyzer intended for the in-vitro quantitative determination of analytes in body fluids.
The Tina-quant Hemoglobin A1cDx Gen.3 assay is intended for use as an aid in diagnosis of diabetes and as an aid in identifying patients who may be at risk for developing diabetes. It is an in vitro diagnostics reagent system intended for quantitative determination of mmol/mol hemoglobin A1c (IFCC) and % hemoglobin A1c (DCCT/NGSP) in hemolysate or whole blood on the Roche/Hitachi cobas c 513 clinical chemistry analyzer. HbA1c determinations are useful for monitoring of long-term blood glucose control in individuals with diabetes mellitus.
The cobas c 513 clinical chemistry analyzer is a fully automated, software controlled analyzer system for in vitro quantitative determination of analytes in human body fluids. The cobas c 513 analyzer system includes a control unit and an analyzer with a closed tube sampling functionality.
The cobas c 513 control unit includes a computer (PC) located in the sampler unit, a touchscreen monitor, soft-keyboard, a mouse (optional use), and a printer. The control unit is used to perform tasks on the analyzer and the PC runs the software that controls the analyzer.
The software manages all instrument functions, all system functions, and all information related to orders and results. The software offers a graphical user interface (GUI) to control all functions by the operator. The control unit contains System Control software including interfaces to a Medical Device Data System (cobas Link) and to the customer Laboratory Information System (LIS).
The analyzer is composed of the sampler unit and the analytical unit. The sampler unit is composed of the rack loading/unloading areas, a barcode reader, a rack rotor, a STAT port, and conveyor lines. It is used to load and unload racks, power on the system, access the PC, and manage the conveyance of samples to the analytical unit.
The analytical unit is comprised of the reagent area, the sample area, and the reaction disk. Samples are conveyed to the analytical unit from the sample unit for photometric analysis before being returned to the sample unit.
Anticoagulated whole blood is hemolyzed either manually or automatically prior to determination of HbAlc by a turbidimetric inhibition immunoassay. Liberated hemoglobin (Hb) in the hemolyzed sample is converted to a derivative having a characteristic absorption spectrum and measured bichromatically. The instrument calculates the % HbAlc from the HbAlc/Hb ratio according to a user selected protocol, either IFCC or NGSP protocols.
The assay offers separate applications that are specific to the sample types whole blood and hemolysate. The Whole Blood Application differs from the Hemolysate Application in the hemolyzing step. For the Whole Blood Application, whole blood samples are placed on the analyzer. Hemolysis occurs onboard the analyzer. For the Hemolysate Application, hemolyzed samples are placed on the analyzer. Hemolysis occurs manually before placing the samples onboard the analyzer. The two applications yield the same results.
This document, a 510(k) premarket notification, describes the performance evaluation of the cobas c 513 Analyzer and cobas c 513 Tina-quant HbA1cDx Gen.3 Assay. It does not describe an AI/ML powered device, but rather an in-vitro diagnostic system for measuring Hemoglobin A1c. Therefore, many of the typical acceptance criteria and study designs relevant to AI/ML based devices (e.g., expert consensus, MRMC studies, training set details) are not applicable or detailed in the same way.
However, I can extract and present the information as per your request based on the provided document, interpreting "acceptance criteria" as the performance criteria the device was tested against and "study" as the non-clinical performance evaluation conducted.
Device Name: cobas c 513 Tina-quant HbA1cDx Gen.3 Assay
Device Type: In-vitro diagnostic reagent system for quantitative determination of HbA1c.
1. Table of Acceptance Criteria and Reported Device Performance
Given that this is a 510(k) for an in-vitro diagnostic (IVD) device, the "acceptance criteria" are not explicitly listed in a single table with pass/fail remarks as might be seen for an AI/ML algorithm. Instead, the document details various performance studies (e.g., precision, linearity, method comparison, interference) and their results, implicitly demonstrating that the device meets an acceptable level of performance for its intended use based on standard IVD guidelines.
The table below summarizes the key performance metrics and their reported values. The "Acceptance Criteria" column reflects typical expected performance ranges or goals for such IVDs based on general industry standards and the context provided by the results themselves. The document states "All acceptance criteria for method comparison were met" and "All acceptance criteria for drug interferences were met," indicating that internal predefined criteria were satisfied.
| Performance Metric | Acceptance Criteria (Implicit/Inferred from Context) | Reported Device Performance (cobas c 513 Tina-quant HbA1cDx Gen.3 Assay) |
|---|---|---|
| Limit of Blank (LoB) | Expected to be very low, indicating minimal signal in the absence of analyte. | Hb: 0.50 g/dL (0.31 mmol/L) HbA1c: 0.19 g/dL (0.12 mmol/L) |
| Limit of Detection (LoD) | Expected to be low, indicating the lowest concentration reliably detected. | Hb: 1.00 g/dL (0.62 mmol/L) HbA1c: 0.29 g/dL (0.18 mmol/L) |
| Precision (Total %CV) (Hemolysate) | Expected to be low across the measuring range (e.g., typically < 5-10% depending on concentration). | Range across human samples (4.63% to 13.8% HbA1c): 0.9% - 1.3% |
| Precision (Total %CV) (Whole Blood) | Expected to be low across the measuring range (e.g., typically < 5-10% depending on concentration). | Range across human samples (4.60% to 13.9% HbA1c): 0.9% - 1.6% |
| Method Comparison (Mean Bias vs. NGSP TOSOH) | Expected to be close to zero, with narrow confidence intervals, implying good agreement with the reference method. | Whole Blood: -0.016 (95% CI: -0.260 to 0.227) Hemolysate: -0.138 (95% CI: -0.429 to 0.153) |
| Method Comparison (% Relative Bias) | Expected to be low (e.g., within a few percentage points of the reference). | At various %HbA1c levels: Whole Blood: -0.8% to 0.1% Hemolysate: -2.8% to -0.8% |
| Total Error (TE) | Expected to be within clinically acceptable limits for HbA1c measurement. | Whole Blood: 2.1% - 3.9% Hemolysate: 3.7% - 5.2% |
| Linearity (Pearson's r) | Expected to be very close to 1, indicating a strong linear relationship between measured and theoretical values. | Hb: 0.9999 HbA1c: 0.9990 |
| Measuring Range (Claimed) | Established range should be supported by linearity and other performance data. | Hemoglobin: 4.0 - 40 g/dL (2.48-24.8 mmol/L) HbA1c: 0.3 - 1.93 g/dL (0.186-1.20 mmol/L) This corresponds to 4.2-15.5 % HbA1c (DCCT/NGSP) |
| Matrix Comparison | Mean differences and 95% CIs should be clinically insignificant across different anticoagulants and fill volumes. | Mean Differences: -0.0559 to 0.0393 (within 95% CIs of approx. +/-0.5) |
| Endogenous Interference | No significant interference (defined as >7% deviation in HbA1c) at specified maximum concentrations. | Met criteria; specific maximum concentrations without interference provided for: Lipemia, Bilirubin, Ditaurobilirubin, Glucose, Rheumatoid Factor |
| Drug Interferences | No significant interference (defined as > ±7% deviation) at specified drug concentrations. | Met criteria for all 16 tested drugs. |
| Cross-Reactivity | No significant interference due to common hemoglobin variants and other related substances. | No significant interference with tested Hb variants (HbS, HbC, HbE, HbD, HbA2) unless otherwise specified. HbF > 7% may yield lower than expected values. |
2. Sample Sizes Used for the Test Set and Data Provenance
- Limit of Blank (LoB): One analyte-free sample measured 10-fold with three lots, over six runs distributed over >3 days on one c 513 analyzer. Total 60 measurements per lot.
- Limit of Detection (LoD): Five samples with low-analyte concentration measured 2-fold with three lots, over six runs distributed over >3 days on one c 513 analyzer. Total 60 measurements per lot.
- Precision: Eight samples (two controls, seven human samples with varying HbA1c concentrations) measured in two aliquots per sample, once each in two runs per day for 21 days, on three cobas c 513 analyzers using 3 reagent lots per system.
- Method Comparison: 155 samples (Hemolysate) and 154 samples (Whole Blood) from the secondary NGSP reference laboratory. Samples were tested over a 3-day period with one lot of reagent on one cobas c 513 analyzer.
- Linearity: Two separate dilution series consisting of at least 11 levels, using human hemolysate sample pools. Samples measured in triplicate.
- Matrix Comparison: At least 40 samples of each sample type (anticoagulant and fill volume) and at least 40 half-filled tubes of each sample type. Overall sample count not explicitly summed but implies a substantial number.
- Endogenous Interference: 12 spiked samples (2 HbA1c levels x 6 interferents). Each treated pool (spiked and control) had a dilution series of at least 10 levels. Dilution series tested 10-fold.
- Drug Interferences: Native patient samples at 2 different HbA1c levels (approx. 6% and 8% HbA1c) were spiked with 16 common drugs at two defined concentrations. Samples measured in 10-fold.
- Cross-reactivity: Multiple experiments for each cross-reactant (HbA0, HbA1a+b, Acetylated Hb, Carbamylated Hb, Glycated Albumin, Labile HbA1c). Each involved a series of dilutions or concentration variations, with 10 replicates per sample.
- Hemoglobin Variants: 20 samples each for HbS, HbC, HbE, HbD, Elevated F. 13 samples for HbA2. Each tested once in at least one run.
Data Provenance: The document does not explicitly state the country of origin for the human samples used in these studies. The studies were non-clinical performance evaluations (laboratory-based testing). The samples for method comparison were obtained from a secondary NGSP (National Glycohemoglobin Standardization Program) reference laboratory. This implies a standardized and presumably well-characterized dataset. The studies are prospective in the sense that they are designed performance evaluations conducted specifically for device submission, rather than analysis of existing stored data.
3. Number of Experts Used to Establish the Ground Truth for the Test Set and Their Qualifications
This document describes a laboratory diagnostic device, not an AI/ML algorithm that interprets medical images or other complex data requiring human expert consensus for ground truth.
- Ground truth for HbA1c values: For the method comparison study, the ground truth was established by a secondary NGSP reference laboratory using a Tosoh HPLC system. This is a recognized and standardized reference method for measuring HbA1c. The document mentions "Traceability: This method has been standardized against the approved IFCC reference method for the measurement of HbA1c in human blood and can be transferred to results traceable to DCCT/NGSP by calculation." This indicates that the ground truth is established via an internationally recognized and highly accurate analytical method, rather than through expert human assessment.
Therefore, the concept of "number of experts" and their "qualifications" for establishing ground truth in the context of an AI/ML diagnostic for image interpretation is not directly applicable here.
4. Adjudication Method for the Test Set
Not applicable. As described above, the ground truth for this IVD device is established through a laboratory reference method (Tosoh HPLC system) traceable to international standards (IFCC, NGSP), not through human expert assessment needing adjudication.
5. If a Multi Reader Multi Case (MRMC) Comparative Effectiveness Study Was Done
No. This document describes the performance of an in-vitro diagnostic instrument and assay, not an AI/ML application intended to assist human readers (e.g., radiologists) in interpreting medical images. Therefore, an MRMC study and the concept of "how much human readers improve with AI vs. without AI assistance" are not relevant to this device.
6. If a Standalone (i.e. algorithm only without human-in-the loop performance) Was Done
Yes, in essence. The performance studies detailed in this document (precision, method comparison, linearity, interference, etc.) directly evaluate the cobas c 513 Tina-quant HbA1cDx Gen.3 Assay and cobas c 513 Analyzer as a standalone system for quantitative determination of HbA1c in patient samples. The results are generated entirely by the instrument and reagents, independent of human interpretive input beyond standard laboratory procedures for operating the device and reporting results. There is no "human-in-the-loop" interpretive component that would require a separate "standalone" versus "human-in-the-loop" performance evaluation as might be seen for an AI algorithm.
7. The Type of Ground Truth Used
The ground truth for the HbA1c values, particularly in the method comparison study, was established using a Tosoh HPLC system operated by a secondary NGSP (National Glycohemoglobin Standardization Program) reference laboratory. This method is traceable to the approved IFCC (International Federation of Clinical Chemistry and Laboratory Medicine) reference method and linked to DCCT/NGSP standards. This represents a highly standardized and analytically robust reference method/pathology/analytical outcome data rather than expert consensus on subjective findings.
8. The Sample Size for the Training Set
This document does not describe the development of an AI/ML algorithm that would have separate "training" and "test" sets in the conventional sense. This is an IVD device where performance characteristics are assessed on "test sets" (i.e., various sample groups used in the performance studies) to demonstrate its analytical capability. Therefore, the concept of a "training set" is not applicable here.
9. How the Ground Truth for the Training Set Was Established
Not applicable, as there is no "training set" in the context of an AI/ML algorithm.
{0}------------------------------------------------
Image /page/0/Picture/1 description: The image is a black and white logo for the Department of Health & Human Services - USA. The logo features a stylized image of three human profiles facing to the right, stacked on top of each other. The profiles are connected and form a single, abstract shape. The text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" is arranged in a circular pattern around the image.
Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002
December 19, 2016
ROCHE DIAGNOSTICS OPERATIONS DAVID TRIBBETT REGULATORY PROGRAM MANAGER 9115 HAGUE ROAD INDIANAPOLIS, IN 46250
Re: K160571 Trade/Device Name: cobas c 513 Analyzer, cobas c 513 Tina-quant HbA1cDx Gen.3 Assay Regulation Number: 21 CFR 862.1373 Regulation Name: Hemoglobin Alc test system Regulatory Class: II Product Code: PDJ, JJE Dated: December 08, 2016 Received: December 09, 2016
Dear David Tribbett:
We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.
If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.
Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (OS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.
{1}------------------------------------------------
If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 809), please contact the Division of Industry and Consumer Education at its toll-free number (800) 638 2041 or (301) 796-7100 or at its Internet address
http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to
http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.
You may obtain other general information on your responsibilities under the Act from the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address
http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.
Sincerely yours,
Courtney
Courtney H. Lias, Ph.D. Director Division of Chemistry and Toxicology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health
Enclosure
{2}------------------------------------------------
Indications for Use
510(k) Number (if known) K160571
Device Name cobas c 513 Analyzer cobas c 513 Tina-quant HbA1cDx Gen.3 Assay
Indications for Use (Describe)
The cobas c 513 clinical chemistry analyzer is a fully automated, standalone clinical chemistry analyzer intended for the in-vitro quantitative determination of analytes in body fluids.
The Tina-quant Hemoglobin A1cDx Gen.3 assay is intended for use as an aid in diagnosis of diabetes and as an aid in identifying patients who may be at risk for developing diabetes. It is an in vitro diagnostics reagent system intended for quantitative determination of mmol/mol hemoglobin Alc (IFCC) and % hemoglobin A1c (DCCT/NGSP) in hemolysate or whole blood on the Roche/Hitachi cobas c 513 clinical chemistry analyzer. HbA1c determinations are useful for monitoring of long-term blood glucose control in individuals with diabetes mellitus.
Type of Use (Select one or both, as applicable)
| Registration Use (Part 81 CFR 801 Subpart D) | |
|---|---|
| Over-The-Counter Use |
|X Prescription Use (Part 21 CFR 801 Subpart D)
| | Over-The-Counter Use (21 CFR 801 Subpart C)
CONTINUE ON A SEPARATE PAGE IF NEEDED.
This section applies only to requirements of the Paperwork Reduction Act of 1995.
DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.
The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to:
Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff@fda.hhs.gov
"An agency may not conduct or sponsor, and a person is not required to respond to, a collection of information unless it displays a currently valid OMB number."
{3}------------------------------------------------
cobas c 513 Tina-quant Hemoglobin A1cDx Gen.3 assay 510(k) Summary
This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of 21 CFR 807.92.
| Submitter Name | Roche Diagnostics Operations (RDO) |
|---|---|
| Address | 9115 Hague RoadIndianapolis, IN, 46250, USA |
| Contact | David TribbettPhone: (317)-521-2964FAX: (317)-521-2324Email: david.tribbett@roche.com |
| Date Prepared | January 31, 2016 |
| Proprietary Names | cobas c 513 clinical chemistry analyzerTina-quant Hemoglobin A1cDx Gen.3 |
| Common Names | Analyzer, Chemistry (Photometric, Discrete), for clinical useGlycosylated Hemoglobin Assay |
| Classification Name | Discrete photometric chemistry analyzer for clinical useHemoglobin A1c test system |
| Product Codes | JJE, 21 CFR 862.2160PDJ, 21 CFR 862.1373 |
| Predicate Device | COBAS INTEGRA 800 Tina-quant HbA1cDx Gen.2 assay, K121291 |
| Establishment Registration | 1823260, Roche Diagnostics Corporation |
{4}------------------------------------------------
1. DEVICE DESCRIPTION
Roche Diagnostics plans to introduce the cobas c 513 clinical chemistry analyzer, a new member of the Roche / Hitachi family of analyzers. It has been developed jointly by Roche Diagnostics and Hitachi High Technologies (HHT). It is a standalone, high volume Roche/Hitachi cobas c analyzer, that is to be used with the Tina-quant HbA1cDx Gen.3 Assay which is intended for use with human whole blood and hemolysate samples for monitoring of long term blood glucose control in individuals with diabetes mellitus and as "an aid in diagnosis of diabetes and as an aid in identifying patients who may be at risk for developing diabetes".
Analyzer
The cobas c 513 analyzer is a fully automated, software controlled analyzer system for in vitro quantitative determination of analytes in human body fluids. The cobas c 513 analyzer system includes a control unit and an analyzer with a closed tube sampling functionality.
The cobas c 513 control unit includes a computer (PC) located in the sampler unit, a touchscreen monitor, soft-keyboard, a mouse (optional use), and a printer. The control unit is used to perform tasks on the analyzer and the PC runs the software that controls the analyzer.
The software manages all instrument functions, all system functions, and all information related to orders and results. The software offers a graphical user interface (GUI) to control all functions by the operator. The control unit contains System Control software including interfaces to a Medical Device Data System (cobas Link) and to the customer Laboratory Information System (LIS).
The analyzer is composed of the sampler unit and the analytical unit. The sampler unit is composed of the rack loading/unloading areas, a barcode reader, a rack rotor, a STAT port, and conveyor lines. It is used to load and unload racks, power on the system, access the PC, and manage the conveyance of samples to the analytical unit.
The analytical unit is comprised of the reagent area, the sample area, and the reaction disk. Samples are conveyed to the analytical unit from the sample unit for photometric analysis before being returned to the sample unit.
{5}------------------------------------------------
Assay
Anticoagulated whole blood is hemolyzed either manually or automatically prior to determination of HbAlc by a turbidimetric inhibition immunoassay. Liberated hemoglobin (Hb) in the hemolyzed sample is converted to a derivative having a characteristic absorption spectrum and measured bichromatically. The instrument calculates the % HbAlc from the HbAlc/Hb ratio according to a user selected protocol, either IFCC or NGSP protocols.
The assay offers separate applications that are specific to the sample types whole blood and hemolysate. The Whole Blood Application differs from the Hemolysate Application in the hemolyzing step. For the Whole Blood Application, whole blood samples are placed on the analyzer. Hemolysis occurs onboard the analyzer. For the Hemolysate Application, hemolyzed samples are placed on the analyzer. Hemolysis occurs manually before placing the samples onboard the analyzer. The two applications yield the same results.
Test principle
This method uses tetradecyltrimethyl-ammonium bromide (TTAB) as the detergent in the hemolyzing reagent to eliminate interference from leukocytes (TTAB does not lyse leukocytes). Hemolyzing reagent is part of the test system and is either placed on board the analyzer for the whole blood application or used manually for the hemolysate application. Sample pretreatment to remove labile HbA1c is not necessary. All hemoglobin variants which are glycated at the ßchain N-terminus and which have antibody-recognizable regions identical to that of HbA1c are determined by this assay. Consequently, the metabolic state of patients having uremia or the most frequent hemoglobinopathies (HbAS, HbAC, HbAE, HbAD) can be determined using this assay.
Hemoglobin A1c
The HbA1c determination is based on the turbidimetric inhibition immunoassay (TINIA) for hemolyzed whole blood.
{6}------------------------------------------------
Sample and addition of R1 (buffer/antibody):
Glycohemoglobin (HbA1c) in the sample reacts with anti-HbA1c antibody to form soluble antigen-antibody complexes. Since the specific HbA1c antibody site is present only once on the HbA1c molecule, formation of insoluble complexes does not take place.
Addition of R2 (buffer/polyhapten) and start of reaction:
The polyhaptens react with excess anti-HbA1c antibodies to form an insoluble antibodypolyhapten complex which can be determined turbidimetrically.
Hemoglobin
Liberated hemoglobin in the hemolyzed sample is converted to a derivative having a characteristic absorption spectrum which is measured bichromatically during the preincubation phase (sample + R1) of the above immunological reaction. A separate Hb reagent is consequently not necessary.
The final result is expressed as mmol/mol HbA1c or % HbA1c and is calculated from the HbA1c/Hb ratio as follows:
Protocol 1 (mmol/mol HbA1c acc. to IFCC): HbA1c (mmol/mol) = (HbA1c/Hb) × 1000
Protocol 2 (% HbA1c acc. to DCCT/NGSP): HbA1c (%) = (HbA1c/Hb) × 91.5 + 2.15
Standardization
Traceability: This method has been standardized against the approved IFCC reference method for the measurement of HbA1c in human blood and can be transferred to results traceable to DCCT/NGSP by calculation.
2. INDICATIONS FOR USE
The cobas c 513 analyzer is a fully automated, standalone clinical chemistry analyzer intended for the in-vitro quantitative determination of analytes in body fluids.
The Tina-quant Hemoglobin A1cDx Gen.3 assay is intended for use as an aid in diagnosis of diabetes and as an aid in identifying patients who may be at risk for developing diabetes. It is an
{7}------------------------------------------------
in vitro diagnostics reagent system intended for quantitative determination of mmol/mol hemoglobin A1c (IFCC) and % hemoglobin A1c (DCCT/NGSP) in hemolysate or whole blood on the Roche/Hitachi cobas c 513 clinical chemistry analyzer. HbA1c determinations are useful for monitoring of long-term blood glucose control in individuals with diabetes mellitus.
3. TECHNOLOGICAL CHARACTERISTICS
As the cobas c 513 analyzer is a new member of the Roche/Hitachi clinical chemistry instruments family, a comparison to the cobas c 501 analyzer is shown in the table below to compare the analyzer to other members of the Roche/Hitachi Clinical Chemistry instrument family.
| Topic | cobas c 501 | cobas c 513 |
|---|---|---|
| Basic features | ||
| Intended Use | Fully automated clinical chemistry analyzer intended for the in vitro quantitative / qualitative determination of analytes in body fluids | The cobas c 513 is a fully automated, discrete clinical chemistry analyzer intended for the in-vitro quantitative determination of analytes in body fluids. |
| Measurement principle | Absorbance Photometry (enzymes, substrates, proteins, DAT, TDM) | Absorbance Photometry (enzymes, substrates, proteins, DAT, TDM) |
| ISE Potentiometric (electrolytes)Endpoint, Kinetic, | N/A | |
| Reaction modes | Potentiometric | Same |
| N/A | ||
| Throughput Photometry | Max 600 tests per hour without ISE | 400 tests per hour |
| Throughput ISE Potentiometry | Max 600 tests per hour | N/A |
| Analyzer | Standalone module or multiple modules linked | Standalone analyzer |
| Sample Handling | ||
| Typical sample volumes | 1.0 – 35 μL | S1: 1.5-25 μlS2: 1.5-6 μl |
| Sample types | Serum, plasma, urine, CSF and other depending on the chemistry test | Hemolysate, whole blood, Serum/Plasma, Urine |
| Sample handling system | Input of samples via core input buffer using universal sample racks | Closed Tube Sampling (CTS) on specified 5 position Racks ( CTS Rack) and universal sample racks |
| Sample capacity On board | 150 | Same |
| Sample identification | Barcode | Same |
| Topic | cobas c 501 | cobas c 513 |
| Reagent Handling | ||
| Reagent volume | 5 – 180 μL | 15-150 μl |
| Reagent container(electrolytes) | Plastic bottles open | N/A |
| Reagent container(non-electrolytes) | Plastic bottles closed via pierceable screwcaps | Plastic bottles closed via screwcapsThe cap of the reagent bottle isdecapped (opened) by operator |
| Reagent access | Reagent cassette caps pierced onboard by theinstrument | Cassette caps to be opened beforeplacing on instrument |
| Onboard storageTemperature | Refrigerated5 -12 °C | Refrigerated5-15°C |
| Reagent bottle / Cassetteidentification | Barcode | RFID |
| On board reagent storagecapacity | 60 rotor channels on 1 rotor to run 60 reagent kitsin parallel | Same |
| System cycle time | 6 sec | 7.2 sec |
| Reagent mixing | Ultrasonic | Same |
| Auto rerun | Available | Same |
| Application informationtransfer to instrument | Via remote transfer or CD | Via remote transfer |
| Pipetting System | ||
| Sample and reagentSyringes | XY robotic | Same |
| Reagent probes | 2 polished steel probes | Same |
| Sample probes | 1 polished steel probe | 2 polished steel probes |
| Probe cleaning | Automatic for all probes | Same |
| Liquid level detection (LLD)sample | Electrostatic | Electrostatic for open tubes (S1), noLLD for CTS (S2) |
| Liquid level detectionreagent | Initial Cassette Volume Check (ICVC) | Electrostatic (Capacitance) |
| Clot detection | Provided | Same |
| Test Reaction Chamber | ||
| Temperature control | Circulating water bath at 37°C | Same |
| Cuvettes | Multiple use | Same |
| Detection Information | ||
| Spectrophotometer | Gradient photometer with discrete photodiodes infixed array | Same |
| Light source | Tungsten/halogen | Same |
| Light path | 5.6 mm | 5 mm |
| Measuring unit | 1 | Same |
| Wavelengths | 340,376,415,450,480,505,546, 570, 600,660,700,800 nm | Same |
| Calibration and Quality Control | ||
| Calibrators | Multiple use | Same |
| Calibration modes | Linear, nonlinear | Same |
| Calibration Stability | Typically each lot or 12 weeks for same reagenton board | For HbA1c: 4 weeks during shelf live |
| Online QC | Yes | No automatic QC, but QC timeoutfunction |
| Control storage oninstrument | In remote buffer at ambient temperature | none |
| Calibrator/control valuetransfer | Via remote transfer or CD | Via remote transfer |
| Topic | cobas c 501 | cobas c 513 |
| Internal qualitymanagement system | Available to monitor and validate test results | Same |
| Software and Interfaces | ||
| Host Interface | RS232C bi-directional | LAN/Ethernet |
| Printer | Laser | Any printer supported by the OS |
| Display | Keyboard + touch-screenOptional use of mouse | Keyboard + touch-screen: soft-keyboard; Optional use of mouse |
| Software | cobas 6000 modular system software | c 513 instrument software |
| Configuration | One or several analytical units with one PC andone Core | One stand-alone analytical unit withone PC |
| Units controlled | cobas c501 and cobas e601 | 513 analytical unit, one unit |
| Functions performed | Data input, sample processing, result calculation,result reporting, quality control | Same |
| PC (Controller Unit)functions | Data input (keyboard, disc),Data output (screen, printer) | Data input (keyboard, disc)no input from USB,DVD-RAMData output (screen, printer + USB DVD-RAM) |
| Core Unit functions | Realtime database, data input and output ( viaHOST Communication ) control of sample conveyer | Same |
| Analytical Unit(s) function | Control of analytic processes (pipetting, incubation,detection) Primary Signal processing | Same |
| Data storage | Real time database in Core unit(storage of System and Application parameters,Calibration Data ,QC Data, Sample results, Alarmhistory) | Same |
| Software-controlled testcountdown | Available | Same |
| Result calculation | Automated measuring of signal for kinetic andendpoint methods according to cycle time andautomated calculation of concentrations viacalibration curve | Same |
| User management | Yes | Same |
| Flagging of errors | Available | Same |
Substantial Equivalence – Analyzer Similarities
{8}------------------------------------------------
{9}------------------------------------------------
Substantial Equivalence – Assay Similarities
While the c 513 analyzer is a new member of the Roche /Hitachi family of analyzers, none of the analyzers in this family have received 510(k) clearance for an HbA1c "aid in diagnosis" assay for both human whole blood and hemolysate sample types. Sample type is a key element of the intended use, which necessitates the COBAS Integra 800 Tina-quant Hemoglobin A1cDx Gen.2 to be the predicate device since it has been cleared for aid in diagnosis for both of these sample types.
{10}------------------------------------------------
| Feature | Predicate Device: COBAS INTEGRA 800Tina-quant HbA1cDx Gen.2 assayK121291 | Candidate Device: cobas c 513 Tina-quant HbA1cDx Gen.3 assay |
|---|---|---|
| Intended Use | This test is to be used as an aid indiagnosis of diabetes and as an aid inidentifying patients who may be at risk fordeveloping diabetes. | This test is intended for use as an aid indiagnosis of diabetes and as an aid inidentifying patients who may be at risk fordeveloping diabetes. It is an in vitrodiagnostics reagent system intended forquantitative determination of mmol/molhemoglobin A1c (IFCC) and %hemoglobin A1c (DCCT/NGSP) inhemolysate or whole blood on theRoche/Hitachi cobas c 513 clinicalchemistry analyzer. HbA1cdeterminations are useful for monitoring oflong-term blood glucose control inindividuals with diabetes mellitus. |
| Sample Types | Anticoagulated venous or capillary bloodAcceptable anticoagulants for both thehemolysate and whole blood applicationsinclude:Li-HeparinK2-EDTAK3-EDTANa-HeparinNaF/K-OxalateNaF/Na2-EDTA | Anticoagulated venous bloodAcceptable anticoagulants for both thehemolysate and whole blood applicationsinclude:Li-HeparinK2-EDTAK3-EDTAEDTA/Fluoride |
| Instrument Platform | COBAS Integra 800• Absorbance Photometry | Cobas c 513• Same |
| Calibrator | Cfas HbA1c | Same |
| Calibration Frequency | Each lot, every 29 days, and as requiredfollowing quality control procedures | 29 days |
| Calibration Mode | Logit/log 5 | Hb: linearHbA1c : RCM4 |
| Controls | PreciControl HbA1c norm and path | Same |
| Reagent Stability | Unopened:• 2-8°C until expiration dateOn-bard in use:• 2-8°C for 28 days | Same |
| Reporting Units | % HbA1c NGSP/DCCT and mmol/molIFCC | Same |
| Antibody | Polyclonal anti-HbA1c from sheep blood | Same |
| Feature | Predicate Device: COBAS INTEGRA 800Tina-quant HbA1cDx Gen.2 assayK121291 | Candidate Device: cobas c 513 Tina-quant HbA1cDx Gen.3 assay |
| Test Principle | The anticoagulated whole blood specimenis hemolyzed automatically on the COBASINTEGRA 800 analyzers with COBASINTEGRA Hemolyzing Reagent Gen.2.This method uses TTAB as the detergentin the hemolyzing reagent to eliminateinterference from leukocytes (TTAB doesnot lyse leukocytes). Sample pretreatmentto remove labile HbA1c is not necessary.All hemoglobin variants which are glycatedat the β-chain N-terminus and which haveantibody-recognizable regions identical tothat of HbA1c are determined by thisassay. Consequently, the metabolic stateof diabetic patients having uremia or themost frequent hemoglobinopathies (HbAS,HbAC, HbAE, HbAD) can be determinedby this assay. | This method uses tetradecyltrimethyl-ammonium bromide (TTAB) as thedetergent in the hemolyzing reagent toeliminate interference from leukocytes(TTAB does not lyse leukocytes). Samplepretreatment to remove labile HbA1c isnot necessary.All hemoglobin variants which areglycated at the β-chain N-terminus andwhich have antibody-recognizable regionsidentical to that of HbA1c are determinedby this assay. Consequently, themetabolic state of patients having uremiaor the most frequent hemoglobinopathies(HbAS, HbAC, HbAE, HbAD) can bedetermined using this assay. |
| Determination of HbA1c | Turbidimetric immunoinhibition (TINIA).Antigen-antibody complexes are formedand excess Ab aggregate with polyhaptento form insoluble complexes | HbA1c determination is based on theturbidimetric inhibition immunoassay(TINIA) for hemolyzed whole blood.Glycohemoglobin in the sample reactswith anti-HbA1c to form soluble antigen-antibody complexes. Polyhaptens reactwith excess anti-HbA1c to form aninsoluble antibody-polyhapten complexwhich can be measured turbidimetrically. |
| Determination of Hb | Bichromatic photometric determinationafter conversion to a colored derivate | Liberated hemoglobin in the hemolyzedsample is converted to a derivative havinga characteristic absorption spectrumwhich is measured bichromatically. |
| Determination of % HbA1c | The final result is expressed as % HbA1cand is calculated from the HbA1c/Hb ratioper DCCT/NGSP as follows:HbA1c (%) = (HbA1c/Hb) x 91.5 + 2.15 | Same |
| Claimed Measuring Range | Hemoglobin:● 4-35 g/dL (2.48 – 21.7 mmol/L)HbA1c:● 0.3-3.4 g/dL (0.186-2.11 mmol/L) | Hemoglobin:● 4-40 g/dL (2.48-24.8 mmol/L)HbA1c:● 0.3-1.93 g/dL (0.186-1.2 mmol/L) |
| Traceability | The assigned HbA1c and total hemoglobinvalues of the cobas c Tina-quantHemoglobin A1c Gen.3 assay is certifiedwith the National GlycohemoglobinStandardization Program (NGSP). TheNGSP certification expires in one year. | Same |
| Feature | Predicate Device: COBAS INTEGRA 800Tina-quant HbA1cDx Gen.2 assayK121291 | Candidate Device: cobas c 513 Tina-quant HbA1cDx Gen.3 assay |
| Reagent Composition | R1 Antibody Reagent:MES(2-morpholinoethane sulfonic acid) buffer: 0.025 mol/L TRIS (Tris(hydroxymethyl) aminomethane) buffer: 0.015 mol/L, pH 6.2 HbA1c antibody (ovine serum): ≥ 0.5 mg/ml detergents; stabilizers; preservatives SR Polyhapten Reagent: MES buffer: 0.025 mol/L | R1 Antibody Reagent:MES(2-morpholinoethane sulfonic acid) buffer: 0.025 mol/L TRIS (Tris(hydroxymethyl) aminomethane) buffer: 0.015 mol/L, pH 6.2 HbA1c antibody (ovine serum): ≥ 0.5 mg/ml detergents; stabilizers; preservatives R3 Polyhapten Reagent: MES buffer: 0.025 mol/L |
| TRIS buffer: 0.015 mol/L, pH 6.2 HbA1c polyhapten: > 8µg/mL stabilizers; preservatives | TRIS buffer: 0.015 mol/L, pH 6.2 HbA1c polyhapten: > 8µg/mL stabilizers; preservatives | |
| A1CD (Hemolyzing Reagent): Aqueous buffered matrix, pH 7.25 Tetradecyltrimethylammonium bromide: 36 g/L sodium dihydrogenphosphate monohydrate: 16 mmol/L | A1CD (Hemolyzing Reagent): Aqueous buffered matrix, pH 7.25 Tetradecyltrimethylammonium bromide: 36 g/L sodium dihydrogenphosphate monohydrate: 16 mmol/L | |
| sodium monohydrogenphosphate dihydrate: 64 mmol/L stabilizers; preservatives | sodium monohydrogenphosphate dihydrate: 64 mmol/L stabilizers; preservatives |
{11}------------------------------------------------
{12}------------------------------------------------
{13}------------------------------------------------
4. NON-CLINICAL PERFORMANCE EVALUATION
Performance characteristics were evaluated with the Tina-quant HbA1c Gen.3 reagent on the cobas c 513 analyzer.
This assay offers two sample type-specific applications, one for manually hemolysed samples and one for whole blood samples; thus they are named the Hemolysate and Whole Blood Applications.
The Tina-quant HbA1c Gen.3 assay first measures total hemoglobin (Hb) and glycated hemoglobin (HbA1c) in terms of mmol/L. Then the analyzer calculates the HbA1c/Hb ratio according to the corresponding protocol, either IFCC or DCCT/NGSP. IFCC protocol reports the ratio in terms of mmol/mol HbA1c while the DCCT/NGSP protocol reports the ratio in terms of % HbA1c. Performance characteristics that support the measuring ranges claimed for Hb and HbA1c include limit of detection and linearity. They report results in terms of Hb and HbA1c individually. A patient sample value is reported in terms of the ratio of glycated to total hemoglobin. Method comparison, control recovery, and precision are evaluated in terms of the ratio. The protocol according to DCCT/NGSP was chosen for these studies.
The following performance data were provided in support of the substantial equivalence determination:
4.1. Limit of Blank and Limit of Detection (LoB and LoD)
4.1.1. Limit of Blank
For determination of LoB one analyte free sample will be measured with three lots in 10-fold determination. Six runs distributed over > 3 days using one c 513 analyzer will be performed. In total, 60 measurements will be obtained per lot. Data analysis will be based on determination of the 95th percentile of the 60 measured values. In our design (n=60) the 95th percentile is the average of the 57th and 58th value (see also EP17-A2).
{14}------------------------------------------------
Limit of Detection 4.1.2.
For determination of LoD, five samples with low-analyte concentration will be measured with three lots in two-fold determination. Six runs distributed over > three days on one c 513 analyzer will be performed. In total, 60 measurements will be obtained per lot.
LoD is defined as the concentration, at which there is a 95% probability that a sample contains analyte.
| Result (g/dL) | Result (mmol/L) | ||
|---|---|---|---|
| Limit of Blank | Hb | 0.50 | 0.31 |
| HbA1c | 0.19 | 0.12 | |
| Limit of Detection | Hb | 1.00 | 0.62 |
| HbA1c | 0.29 | 0.18 |
LoB and LoD Results
4.2. Precision
Precision experiments were performed in Accordance with CLSI Guideline EP5-A2 to evaluate repeatability (within-run precision) and intermediate precision of within-laboratory precision (total precision).
Two aliquots per sample were measured once each in two runs per day for 21 days on three cobas c 513 analyzers using 3 reagent lots per system. Eight total samples were evaluated in each run: two controls, PreciControl HbA1c norm and path, and 7 human samples with HbAc concentrations near 5%, 6.5%, 7.0%, 8.0%, 10.5%, 12%, and 14%.
The samples were randomized in each run separately. The data set was completed for the 21 days. For each sample, the following was calculated: mean, repeatability and intermediate precision as CV and SD values, and the upper 95% confidence interval for SD and CV values.
{15}------------------------------------------------
| c513 (Hemolysate) - instrument 1 | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Mean | Repeatability | Between-run | Between-day | Between-lot | Total | |||||
| %HbA1c | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| Human1,4.59% | 0.02 | 0.5 | 0.02 | 0.4 | 0.02 | 0.4 | 0.02 | 0.3 | 0.04 | 0.8 |
| Human2.6.18% | 0.03 | 0.4 | 0.00 | 0.0 | 0.02 | 0.4 | 0.02 | 0.4 | 0.04 | 0.7 |
| Human3.6.97% | 0.03 | 0.4 | 0.01 | 0.1 | 0.02 | 0.4 | 0.03 | 0.5 | 0.05 | 0.7 |
| Human4,8.05% | 0.03 | 0.3 | 0.02 | 0.2 | 0.03 | 0.3 | 0.06 | 0.7 | 0.07 | 0.9 |
| Human5.10.3% | 0.04 | 0.4 | 0.01 | 0.1 | 0.03 | 0.3 | 0.05 | 0.5 | 0.08 | 0.7 |
| Human6.11.8% | 0.05 | 0.5 | 0.05 | 0.4 | 0.05 | 0.4 | 0.07 | 0.6 | 0.11 | 0.9 |
| Human7,13.9% | 0.06 | 0.5 | 0.03 | 0.2 | 0.06 | 0.4 | 0.00 | 0.0 | 0.09 | 0.7 |
| Preci Norm,5.40% | 0.02 | 0.4 | 0.01 | 0.1 | 0.02 | 0.4 | 0.01 | 0.2 | 0.03 | 0.6 |
| Preci Path,10.4% | 0.04 | 0.4 | 0.00 | 0.0 | 0.04 | 0.4 | 0.06 | 0.6 | 0.08 | 0.8 |
| c513 (Hemolysate) - instrument 2 | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Mean | Repeatability | Between-run | Between-day | Between-lot | Total | |||||
| %HbA1c | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| Human1,4.64% | 0.02 | 0.5 | 0.02 | 0.5 | 0.02 | 0.4 | 0.01 | 0.3 | 0.04 | 0.02 |
| Human2,6.25% | 0.03 | 0.4 | 0.01 | 0.1 | 0.03 | 0.4 | 0.02 | 0.4 | 0.05 | 0.03 |
| Human3,7.06% | 0.03 | 0.4 | 0.00 | 0.0 | 0.03 | 0.4 | 0.03 | 0.4 | 0.05 | 0.03 |
| Human4,8.14% | 0.03 | 0.4 | 0.00 | 0.0 | 0.03 | 0.4 | 0.04 | 0.4 | 0.06 | 0.03 |
| Human5,10.4% | 0.04 | 0.4 | 0.01 | 0.1 | 0.04 | 0.4 | 0.05 | 0.5 | 0.08 | 0.04 |
| Human6,12.0% | 0.08 | 0.6 | 0.04 | 0.3 | 0.06 | 0.5 | 0.11 | 0.9 | 0.15 | 1.3 |
| Human7,13.8% | 0.05 | 0.4 | 0.04 | 0.3 | 0.04 | 0.3 | 0.07 | 0.5 | 0.10 | 0.05 |
| Preci Norm,5.46% | 0.03 | 0.5 | 0.01 | 0.2 | 0.02 | 0.4 | 0.02 | 0.4 | 0.04 | 0.03 |
| Preci Path,10.5% | 0.04 | 0.4 | 0.02 | 0.2 | 0.04 | 0.4 | 0.05 | 0.5 | 0.08 | 0.04 |
{16}------------------------------------------------
| c513 (Hemolysate) - instrument 3 | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Mean | Repeatability | Between-run | Between-day | Between-lot | Total | |||||
| %HbA1c | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| Human1,4.67% | 0.02 | 0.5 | 0.03 | 0.7 | 0.01 | 0.3 | 0.03 | 0.6 | 0.05 | 1.1 |
| Human2,6.26% | 0.03 | 0.4 | 0.02 | 0.3 | 0.02 | 0.3 | 0.03 | 0.5 | 0.05 | 0.8 |
| Human3,7.06% | 0.03 | 0.4 | 0.03 | 0.4 | 0.02 | 0.2 | 0.04 | 0.6 | 0.06 | 0.9 |
| Human4,8.14% | 0.03 | 0.4 | 0.02 | 0.2 | 0.03 | 0.4 | 0.07 | 0.8 | 0.08 | 1.0 |
| Human5.10.4% | 0.04 | 0.4 | 0.02 | 0.2 | 0.03 | 0.3 | 0.09 | 0.9 | 0.11 | 1.1 |
| Human6.11.8% | 0.06 | 0.5 | 0.03 | 0.2 | 0.04 | 0.4 | 0.09 | 0.7 | 0.12 | 1.0 |
| Human7,13.8% | 0.06 | 0.4 | 0.05 | 0.3 | 0.04 | 0.3 | 0.09 | 0.7 | 0.13 | 0.9 |
| Preci Norm,5.49% | 0.02 | 0.4 | 0.02 | 0.4 | 0.02 | 0.3 | 0.03 | 0.5 | 0.04 | 0.8 |
| Preci Path.10.5% | 0.04 | 0.4 | 0.03 | 0.2 | 0.03 | 0.3 | 0.09 | 0.9 | 0.11 | 1.1 |
| c513 Hemolysate - all 3 instruments | ||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Mean%HbA1c | Repeatability | BetweenRun | BetweenDay | BetweenLot | BetweenInstrument | Total | ||||||
| SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV | |
| Human1,4.63% | 0.02 | 0.5 | 0.03 | 0.6 | 0.02 | 0.4 | 0.02 | 0.4 | 0.04 | 0.9 | 0.06 | 1.3 |
| Human2,6.23% | 0.03 | 0.4 | 0.01 | 0.2 | 0.02 | 0.4 | 0.03 | 0.4 | 0.04 | 0.7 | 0.06 | 1.0 |
| Human3,7.03% | 0.03 | 0.4 | 0.02 | 0.2 | 0.02 | 0.3 | 0.04 | 0.5 | 0.05 | 0.7 | 0.07 | 1.0 |
| Human4,8.11% | 0.03 | 0.4 | 0.01 | 0.2 | 0.03 | 0.4 | 0.06 | 0.7 | 0.04 | 0.5 | 0.08 | 1.0 |
| Human5,10.4% | 0.04 | 0.4 | 0.02 | 0.2 | 0.04 | 0.3 | 0.07 | 0.7 | 0.03 | 0.3 | 0.10 | 0.9 |
| Human6,11.9% | 0.06 | 0.5 | 0.04 | 0.3 | 0.05 | 0.5 | 0.09 | 0.7 | 0.09 | 0.7 | 0.15 | 1.3 |
| Human7,13.8% | 0.06 | 0.4 | 0.04 | 0.3 | 0.05 | 0.4 | 0.07 | 0.5 | 0.02 | 0.1 | 0.11 | 0.8 |
| PreciNorm5.45% | 0.02 | 0.4 | 0.01 | 0.3 | 0.02 | 0.4 | 0.02 | 0.4 | 0.04 | 0.8 | 0.06 | 1.1 |
| Preci Path,10.4% | 0.04 | 0.4 | 0.01 | 0.1 | 0.04 | 0.4 | 0.07 | 0.7 | 0.03 | 0.3 | 0.10 | 0.9 |
{17}------------------------------------------------
| c513 (Whole Blood) - instrument 1 | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Mean%HbA1c | Repeatability | Between-run | Between-day | Between-lot | Total | |||||
| SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV | |
| Human1,4.54% | 0.03 | 0.6 | 0.02 | 0.5 | 0.01 | 0.3 | 0.07 | 1.5 | 0.08 | 1.7 |
| Human2,6.21% | 0.02 | 0.4 | 0.01 | 0.1 | 0.02 | 0.3 | 0.01 | 0.2 | 0.03 | 0.6 |
| Human3,6.97% | 0.03 | 0.4 | 0.01 | 0.2 | 0.02 | 0.3 | 0.01 | 0.2 | 0.04 | 0.6 |
| Human4,8.10% | 0.04 | 0.5 | 0.01 | 0.1 | 0.03 | 0.4 | 0.04 | 0.5 | 0.06 | 0.8 |
| Human5,10.5% | 0.04 | 0.4 | 0.04 | 0.4 | 0.01 | 0.1 | 0.09 | 0.8 | 0.10 | 1.0 |
| Human6,11.8% | 0.05 | 0.4 | 0.04 | 0.4 | 0.08 | 0.6 | 0.10 | 0.8 | 0.14 | 1.2 |
| Human7,13.9% | 0.06 | 0.5 | 0.06 | 0.4 | 0.05 | 0.4 | 0.06 | 0.4 | 0.12 | 0.8 |
| Preci Norm,5.42% | 0.03 | 0.5 | 0.01 | 0.2 | 0.02 | 0.4 | 0.03 | 0.5 | 0.05 | 0.8 |
| Preci Path,10.6% | 0.04 | 0.4 | 0.04 | 0.4 | 0.03 | 0.3 | 0.08 | 0.8 | 0.10 | 1.0 |
| c513 (Whole Blood) - instrument 2 | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Mean | Repeatability | Between-run | Between-day | Between-lot | Total | |||||
| %HbA1c | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| Human1,4.64% | 0.03 | 0.6 | 0.03 | 0.7 | 0.01 | 0.2 | 0.02 | 0.4 | 0.05 | 1.0 |
| Human2,6.29% | 0.03 | 0.4 | 0.01 | 0.2 | 0.02 | 0.3 | 0.02 | 0.3 | 0.04 | 0.6 |
| Human3,7.05% | 0.03 | 0.5 | 0.03 | 0.5 | 0.01 | 0.1 | 0.02 | 0.2 | 0.05 | 0.7 |
| Human4,8.17% | 0.04 | 0.5 | 0.03 | 0.4 | 0.02 | 0.3 | 0.05 | 0.6 | 0.07 | 0.9 |
| Human5.10.5% | 0.05 | 0.5 | 0.03 | 0.3 | 0.02 | 0.2 | 0.07 | 0.6 | 0.09 | 0.8 |
| Human6,12.1% | 0.07 | 0.6 | 0.04 | 0.3 | 0.07 | 0.6 | 0.15 | 1.3 | 0.19 | 1.5 |
| Human7,13.9% | 0.06 | 0.4 | 0.05 | 0.3 | 0.08 | 0.6 | 0.13 | 0.9 | 0.17 | 1.2 |
| Preci Norm,5.51% | 0.03 | 0.5 | 0.02 | 0.4 | 0.01 | 0.2 | 0.01 | 0.2 | 0.04 | 0.7 |
| Preci Path,10.6% | 0.05 | 0.5 | 0.04 | 0.4 | 0.01 | 0.1 | 0.07 | 0.6 | 0.09 | 0.9 |
{18}------------------------------------------------
| c513 (Whole Blood) - instrument3 | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Mean%HbA1c | Repeatability | Between-run | Between-day | Between-lot | Total | |||||
| SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV | |
| Human1,4.61% | 0.03 | 0.7 | 0.02 | 0.4 | 0.02 | 0.3 | 0.01 | 0.2 | 0.04 | 0.9 |
| Human2,6.27% | 0.03 | 0.4 | 0.02 | 0.3 | 0.01 | 0.2 | 0.04 | 0.6 | 0.05 | 0.8 |
| Human3,7.04% | 0.04 | 0.5 | 0.02 | 0.3 | 0.01 | 0.2 | 0.04 | 0.6 | 0.06 | 0.9 |
| Human4,8.21% | 0.04 | 0.5 | 0.04 | 0.4 | 0.01 | 0.1 | 0.09 | 1.1 | 0.10 | 1.2 |
| Human5,10.6% | 0.04 | 0.4 | 0.04 | 0.4 | 0.00 | 0.0 | 0.13 | 1.2 | 0.14 | 1.3 |
| Human6,12.0% | 0.06 | 0.5 | 0.05 | 0.4 | 0.05 | 0.4 | 0.07 | 0.6 | 0.11 | 0.9 |
| Human7,13.8% | 0.07 | 0.5 | 0.03 | 0.2 | 0.04 | 0.3 | 0.27 | 2.0 | 0.28 | 2.1 |
| Preci Norm,5.48% | 0.03 | 0.5 | 0.01 | 0.2 | 0.01 | 0.3 | 0.01 | 0.3 | 0.04 | 0.7 |
| Preci Path,10.7% | 0.05 | 0.5 | 0.06 | 0.6 | 0.00 | 0.0 | 0.13 | 1.2 | 0.15 | 1.4 |
| c513 - whole blood - all 3 instruments | ||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Mean%HbA1c | Repeatability | BetweenRun | BetweenDay | Between Lot | BetweenInstrument | Total | ||||||
| SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV | |
| Human1,4.60% | 0.03 | 0.6 | 0.02 | 0.5 | 0.01 | 0.3 | 0.04 | 0.9 | 0.04 | 1.0 | 0.07 | 1.6 |
| Human2,6.26% | 0.03 | 0.4 | 0.01 | 0.2 | 0.02 | 0.3 | 0.02 | 0.4 | 0.04 | 0.6 | 0.06 | 0.9 |
| Human3,7.02% | 0.03 | 0.5 | 0.02 | 0.4 | 0.02 | 0.2 | 0.03 | 0.4 | 0.04 | 0.5 | 0.06 | 0.9 |
| Human4,8.16% | 0.04 | 0.5 | 0.03 | 0.3 | 0.02 | 0.3 | 0.06 | 0.8 | 0.04 | 0.5 | 0.09 | 1.1 |
| Human5,10.5% | 0.04 | 0.4 | 0.04 | 0.4 | 0.01 | 0.1 | 0.10 | 0.9 | 0.00 | 0.0 | 0.11 | 1.1 |
| Human6,12.0% | 0.06 | 0.5 | 0.04 | 0.3 | 0.07 | 0.6 | 0.11 | 0.9 | 0.11 | 0.9 | 0.19 | 1.5 |
| Human7,13.9% | 0.06 | 0.5 | 0.05 | 0.3 | 0.06 | 0.4 | 0.18 | 1.3 | 0.00 | 0.0 | 0.20 | 1.5 |
| PreciNorm5.47% | 0.03 | 0.5 | 0.01 | 0.3 | 0.02 | 0.3 | 0.02 | 0.3 | 0.04 | 0.8 | 0.06 | 1.1 |
| PreciPath,10.6% | 0.05 | 0.5 | 0.05 | 0.5 | 0.01 | 0.1 | 0.09 | 0.9 | 0.00 | 0.0 | 0.12 | 1.1 |
{19}------------------------------------------------
4.3. Method Comparison
A method comparison study was performed to compare the sample results from the candidate method, Tina-quant HbA1c Gen.3 on the cobas c 513, to results from the TOSOH 8 HPLC. This study will be conducted with both the Tina-quant HbA1c Gen.3 Whole Blood Application and Hemolysate Application.
One hundred and fifty-five (155) samples from the secondary NGSP reference laboratory were used in the evaluation. These samples were measured by the secondary NGSP reference laboratory using a Tosoh HPLC system (X axis) and by the Roche Tina-quant HbA1c Gen.3 test system (Y axis). Samples were tested over a 3 day period with one lot of reagent on one cobas c 513 analyzer.
All acceptance criteria for method comparison were met. The difference plots show there is good agreement between the Roche method and the NGSP TOSOH HPLC reference method. The table below summarizes the bias between the Tina-quant HbA1c Gen.3 (Whole Blood and Hemolysate applications) and the NGSP's TOSOH HPLC reference method.
| Tina-quant HbA1c Gen.3Whole Blood Application | Tina-quant HbA1c Gen.3Hemolysate Application | |
|---|---|---|
| Mean bias vs. NGSP TOSOH | -0.016 | -0.138 |
| Mean bias at lower 95% CI | -0.260 | -0.429 |
| Mean bias at upper 95% CI | 0.227 | 0.153 |
Difference Plot Analysis Data Summary
The table below summarizes the percent relative bias at various points in the measuring range for both applications.
| %HbA1c | Bias limit | % Relative BiasWhole Blood Application | % Relative BiasHemolysate Application |
|---|---|---|---|
| >5% | ≤ 5.0% | 0.1% | -0.8% |
| 6.0% | ≤ 3.5% | -0.2% | -1.4% |
| 6.5% | ≤ 3.5% | -0.3% | -1.6% |
| 7.0% | ≤ 3.5% | -0.4% | -1.8% |
| >12.0% | ≤ 10% | -0.8% | -2.8% |
Bias at Concentration Data Summary
{20}------------------------------------------------
Method Comparison Sample Distribution
| Hemolysate | ||
|---|---|---|
| %HbA1c | SamplesTested | % of Total |
| ≤ 5% | 6 | 3.9% |
| 5 – 6% | 25 | 16.1% |
| 6 – 6.5% | 30 | 19.4% |
| 6.5 – 7% | 36 | 23.2% |
| 7 – 8% | 25 | 16.1% |
| 8 – 9 % | 14 | 9.0% |
| > 9% | 19 | 12.3% |
| Total | 155 | 100.0% |
| Whole blood | ||||
|---|---|---|---|---|
| % | Samples | % of | ||
| HbA1c | Tested | Total | ||
| ≤ 5% | 6 | 3.9% | ||
| 5 - 6% | 26 | 16.9% | ||
| 6 - 6.5% | 30 | 19.5% | ||
| 6.5 – 7% | 36 | 23.4% | ||
| 7-8% | 25 | 16.2% | ||
| 8-9% | 14 | 9.1% | ||
| > 9% | 17 | 11.0% | ||
| Total | 154 | 100.0% |
4.4. Total Error Near the Cut-off
Using the results of bias estimation (%Bias) in the method comparison study and precision estimates in the precision study, the Total Error (TE) at the following concentrations (5%, 6.5%, 8% and 12%) was calculated as follows: %TE =%Bias| + 1.96 * %CV * (1+%Bias). The results are presented in the tables below.
Total Error - Whole Blood Application
| % HbA1c | %BIAS | %CV | Total Error (%) |
|---|---|---|---|
| 5.0% HbA1c | 0.7 | 1.6 | 3.9 |
| 6.5% HbA1c | 0.3 | 0.9 | 2.1 |
| 8.0% HbA1c | 0.1 | 1.1 | 2.3 |
| 12.0% HbA1c | 0.2 | 1.5 | 3.1 |
Total Error – Hemolysate Application
| % HbA1c | %BIAS | %CV | Total Error (%) |
|---|---|---|---|
| 5.0% HbA1c | 1.1 | 1.3 | 3.7 |
| 6.5% HbA1c | 1.7 | 1.0 | 3.7 |
| 8.0% HbA1c | 2.0 | 1.0 | 4.0 |
| 12.0% HbA1c | 2.6 | 1.3 | 5.2 |
{21}------------------------------------------------
4.5. Linearity
Two separate dilution series consisting of at least 11 levels were prepared using the human hemolysate sample pools with either HbA1c or Hb with concentrations above the upper end of the corresponding measuring range. Hemolyzing reagent was used for the diluent. Samples were measured in triplicate and data analysis was done separately for each sample. Linear regression analysis was done according to EP6-A. Additionally, first order regression statistics using theoretical values determined from sample pools with known concentrations is provided in the table below.
| Application | Analyte | Low End of LinearRange | High End of LinearRange | Slope | Intercept | Pearson'sr | ||
|---|---|---|---|---|---|---|---|---|
| g/dL | mmol/L | g/dL | mmol/L | |||||
| Hemolysate | Hb | 3.4 | 2.09 | 40.3 | 25.0 | 0.983 | 0.051 | 0.9999 |
| HbA1c | 0.28 | 0.177 | 2.85 | 1.77 | 0.972 | 0.002 | 0.9990 |
Empirical First Order Linear Regression Results
Claimed Ranges:
Hemoglobin: 4.0 - 40 g/dL (2.48-24.8 mmol/L)
HbA1c: 0.3 - 1.93 g/dL (0.186-1.20 mmol/L)
This corresponds to a measuring range of 23-146 mmol/mol HbA1c (IFCC) and 4.2-15.5 % HbA1c (DCCT/NGSP) at a typical hemoglobin concentration of 8.2 mmol/L.
4.6. Matrix Comparison
A matrix comparison study was performed to evaluate different anticoagulants on analyte recovery. At least 40 samples of each sample type and at least 40 half-filled tubes of each sample type were evaluated. The half-filled (double concentrated) and filled sample tubes were from one donor. Only samples within the limit of icteric, lipemic and hemolytic interference
{22}------------------------------------------------
were used. Samples covered the measuring range. The Hemolysate application was used for these measurements. All acceptance criteria for matrix comparison were met.
| Sample Type | Anticoagulant | Tube Fill | MeanDifference | Upper 95% | Lower 95% |
|---|---|---|---|---|---|
| Hemolysate | K2-EDTA | ½ full | -0.0559 | 0.319 | -0.431 |
| K3-EDTA | Full | 0.000732 | 0.418 | -0.417 | |
| K3-EDTA | ½ full | -0.0351 | 0.506 | -0.576 | |
| Li Heparin | Full | 0.0156 | 0.501 | -0.470 | |
| Li Heparin | ½ full | -0.0285 | 0.504 | -0.561 | |
| NaF/Potox | Full | -0.00732 | 0.459 | -0.473 | |
| NaF/Potox | ½ full | 0.0393 | 0.483 | -0.404 |
Matrix Comparison Results
Endogenous Interference 4.7.
A study evaluated several endogenous substances for potential interference with the measure of % HbA1c. The following six endogenous substances were evaluated:
- Lipemia .
- Bilirubin ●
- . Ditaurobilirubin
- . Glucose
- . Rheumatoid Factor
- Total Protein ●
Pooled whole blood samples with two A1c levels, one near the medical decision level and one above it, were spiked with the maximum level of the above six interferents in separate preparations resulting in 12 spiked samples. These samples were then hemolyzed with Tinaquant HbA1c Hemolyzing Reagent. Another pool, without interferent, was equally hemolyzed. A minimum of a 10-level dilution series was then created for each of the 12 spiked samples by using the interferent free pool as the diluting reagent.
{23}------------------------------------------------
The experiment was performed with one reagent lot, one c 513 analyzer, and in a single run from the same calibration. PreciControl HbA1c norm and path will be the controls. The twelve dilution series were tested ten-fold for % HbA1c using the Hemolysate Application only. Interference is a matter of specificity of the assay which is independent of the application; therefore, interference data on the Hemolysate application will be representative of data on both the Hemolysate and Whole Blood Applications.
The mean of the ten replicates was determined and compared to the result from the reference sample (aliquot with no interfering substance). The comparison was evaluated as a percent deviation. An interferent will be significant if it causes >7% deviation of a measurement in terms of % HbA1c.
Endogenous Interference
| Interferent | Maximum Concentration without interferenceClaimed |
|---|---|
| Lipemia | 600 mg/dL |
| Bilirubin | 60 mg/dL |
| Ditaurobilirubin | 60 mg/dL |
| Glucose | 1000 mg/dL |
| Rheumatoid Factor | 750 IU/mL |
4.8. Drug Interferences
A study evaluated several drugs for potential interference with the measurement of %HbA1c using the hemolysate application. The following drugs were studied:
- . N- Acetylcysteine
- Ampicillin-Na .
- Ascorbic acid .
- Cefoxitin ●
- Heparin ●
- Levodopa ●
- . Methyldopa
{24}------------------------------------------------
- . Metronidazole
- Doxycyclin
- Acetylsalicylic acid ●
- Rifampicin ●
- Cyclosporine ●
- Phenylbutazone ●
- Acetaminophen ●
- Ibuprofen .
- . Theophylline
The 16 commonly used drugs listed above were added to native patient samples and examined for potential effect on % HbA1c determination with the Hemolysate Application. Drug interference testing was performed with hemolysate samples at 2 different HbA1c levels. The two different HbA1c concentrations will be approximately 6% and 8% HbA1c. Each drug was added in two defined concentrations with concentration 1 being several times (typically 5 times) the maximum daily dosage and concentration 2 being the maximum daily dosage level. Concentration 1 was performed for screening purposes only and concentration 2 is the relevant drug concentration for determining interferences with the assay. Samples were measured in tenfold using the cobas c 513 system. The median value was compared to the reference value (HbA1c sample with no drug added) and the deviation from the reference was calculated.
The data was collected using the Hemolysate Application which is acceptable since the assay interference is a matter of specificity of the antibody.
Significant interference is defined as > ±7% deviation from the reference value observed. All acceptance criteria for drug interferences were met.
Cross reactivity 4.9.
Studies were performed to determine if the Tina-quant Hemoglobin A1cDx Gen.3 assay demonstrates cross-reactivity with any of the following hemoglobin fractions and glycated albumin.
{25}------------------------------------------------
- HbA0 ●
- HbA1a+b ●
- Acetylated Hb ●
- Carbamylated Hb ●
- Glycated Albumin .
- Labile HbA1c ●
A series experiments were performed with one reagent lot, one c 513 analyzer, and in a single run from the same calibration. PreciControl HbA1c norm and path function as the controls. Ten replicates of each sample were analyzed. The % recovery of the median for each dilution step related to the median obtained for Pool 1 will be plotted against the dilution step and/or the concentration of the potential cross-reactant.
As with the interference studies, the data was collected using the Hemolysate Application only. This is acceptable because cross-reactivity is a matter of specificity of the assay which is independent of the application. Therefore, cross-reactivity data on the Hemolysate Application are representative of data on both the Hemolysate and Whole Blood Application.
| Cross-Reactant | Max ConcentrationMeasured | Max Concentrationwith no interference | |
|---|---|---|---|
| HbA0 | Level 1 | 120 g/dL | 120 g/dL |
| Level 2 | 120 g/dL | 120 g/dL | |
| HbA1a+b | Level 1 | 1.60 g/dL | 0.96 g/dL |
| Level 2 | 1.60 g/dL | 1.60 g/dL | |
| Acetylated Hb | Level 1 | 2.00 g/dL | 2.00 g/dL |
| Level 2 | 2.00 g/dL | 2.00 g/dL | |
| Carbamylated Hb | Level 1 | 2.0 g/dL | 2.0 g/dL |
| Level 2 | 2.0 g/dL | 2.0 g/dL | |
| Glycated albumin | Level 1 | 10.0 g/dL | 10.0 g/dL |
| Level 2 | 10.0 g/dL | 10.0 g/dL | |
| Labile HbA1c | Level 1 | 1500 mg/dL | 1500 mg/dL |
| Level 2 | 1500 mg/dL | 1500 mg/dL |
Cross-Reactivity
{26}------------------------------------------------
4.10. Hemoglobin Variants
Hemoglobin variant testing was conducted to determine if there was any significant interference with any of the major hemoglobin variants and the Tina-quant Hemoglobin A1cDx Gen.3 assay. Hemoglobin variants are structurally altered hemoglobin molecules with at least one amino acid exchange compared to the normal beta chain of hemoglobin. These changes are caused by mutations in the coding region of the globin genes which encode the protein part of hemoglobin. The most common hemoglobin variants are HbS, HbC, HbD and HbE. Moreover, in some conditions, the fetal hemoglobin HbF is elevated. Also, the erythrocytes of some patients (e.g. beta thalassemia minor) contain elevated levels of HbA2. Therefore, it is crucial to ensure accurate HbA1c results from patients who are carriers of these variants.
| Variant Type | Number of Samples | % Variant | HbA1c % |
|---|---|---|---|
| HbS | 20 | 31 - 42% S | 5.0-14.4 |
| HbC | 20 | 33 - 44% C | 4.7-13.0 |
| HbE | 20 | 27 - 33% E | 5.0-9.7 |
| HbD | 20 | 34 - 42% D | 5.0-10.9 |
| Elevated F | 20 | 2 - 28% F | 5.8-10.1 |
| HbA2 | 13 | 4.3 - 6.2% A2 | 5-10 |
Hemoglobin Variant Samples
Each sample was tested once in at least one run on one c 513 analyzer.
Results obtained with the Tina-quant Hemoglobin A1cDx Gen.3 assay (Hemolysate Application) on the c 513 analyzer will be compared to those obtained with the reference methods.
For purposes of this experiment, the data was collected using the Hemolysate Application. This is acceptable since variant interference is a matter of specificity of the antibody. Therefore, the data shown is representative of both the whole blood and hemolysate applications.
There was no significance interference with any of the major hemoglobin variants. Results are summarized in the table below.
{27}------------------------------------------------
| Percent Relative Bias from Reference Method at Low and High Concentrations of HbA1c Samples | ||||
|---|---|---|---|---|
| ~6.0% HbA1c | ~9.0% HbA1c | |||
| Hb Variant | Relative %Difference | Range | Relative %Difference | Range |
| HbA2 | -2.0% | -3.5 - (-1.3) | 1.2% | -1.2 - 2.5 |
| HbC | -1.3% | -4.0 - 1.79 | -1.3% | -4.79 - 2.45 |
| HbD | -1.5% | -1.9 - 0.8 | -2.3% | -2.4 - (-0.3) |
| HbE | -1.8% | -4.5 - (-0.2) | 0.7% | -0.9 - 2.5 |
| HbS | -1.6% | -4.1 - 3.4 | 2.7% | -0.3 - 3.0 |
| HbF | Specimens containing high amounts of HbF (> 7 %) may yield lower than expected HbA1cvalues. |
Hemoglobin Variant Testing Results
Heterozygous presence of the most common hemoglobin variants (HbAS, HbAD, HbAE) does not interfere. Significant interference was defined as ≥7% change in HbA1c value in the presence of the hemoglobin variant relative to control.
5. CONCLUSIONS
The submitted information in this premarket notification 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.