(146 days)
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 venous whole blood on the cobas c 503 clinical chemistry analyzer. HbA1c determinations are useful for monitoring of long-term blood glucose control in individuals with diabetes mellitus.
Tina-quant Hemoglobin A1cDx Gen.3 assay 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 cobas c 503 clinical chemistry analyzer. 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 and hemolysis occurs onboard the analyzer. For the Hemolysate Application, hemolyzed samples are placed on the analyzer and hemolysis occurs manually before placing the samples onboard the analyzer. The two applications yield the same results. 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. 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.
Here's a breakdown of the acceptance criteria and the study that proves the device meets them, based on the provided FDA 510(k) summary for the Tina-quant Hemoglobin A1cDx Gen.3:
1. A table of acceptance criteria and the reported device performance
The document doesn't explicitly state "acceptance criteria" as a single, consolidated table with pass/fail thresholds. Instead, it presents various performance studies and their results. The implicit acceptance criteria are that the device's performance metrics are acceptable for its intended use and comparable to or better than the predicate device.
Here’s a table summarizing the reported device performance for key metrics:
| Performance Metric | Acceptance Criteria (Implicit) | Reported Device Performance (Tina-quant Hemoglobin A1cDx Gen.3) |
|---|---|---|
| Precision | Acceptable levels of repeatability and intermediate precision for clinical use. | Hemolysate Application: - Reproducibility (total CV%): Most samples/controls show CV% between 1.0% and 1.9%. - Repeatability (CV%): Most samples/controls show CV% between 0.4% and 0.6%. Whole Blood Application: - Reproducibility (total CV%): Most samples/controls show CV% between 1.2% and 1.6%. - Repeatability (CV%): Most samples/controls show CV% between 0.4% and 0.8%. |
| Limit of Blank (LoB) | Detection limit sensitive enough for clinical application. | Hb LoB: 0.0530 mmol/L (0.085 g/dL) HbA1c LoB: 0.0220 mmol/L (0.035 g/dL) This corresponds to 15 mmol/mol (IFCC) and 3.5% HbA1c (DCCT/NGSP) at 13.2 g/dL Hb. |
| Limit of Detection (LoD) | Detection limit sensitive enough for clinical application. | Hb LoD: 0.119 mmol/L (0.192 g/dL) HbA1c LoD: 0.0437 mmol/L (0.07 g/dL) This corresponds to 22 mmol/mol (IFCC) and 4.2% HbA1c (DCCT/NGSP) at 13.2 g/dL Hb. |
| Linearity/Reportable Range | Linear response across the claimed measuring range. | Hemoglobin (Hb): 4 – 40 g/dL (2.48 – 24.8 mmol/L) HbA1c: 0.3 – 2.6 g/dL (0.186 – 1.61 mmol/L) This corresponds to a measuring range of 23-196 mmol/mol HbA1c (IFCC) and 4.2-20.1% HbA1c (DCCT/NGSP). Empirical First Order Regression: Pearson's r for Hb = 0.9999, HbA1c = 0.9990. |
| Endogenous Interferences | No significant interference from common endogenous substances at specified concentrations. | Demonstrated claimed maximum concentrations without interference for: Bilirubin (60 mg/dL), Ditaurobilirubin (60 mg/dL), Lipemia (400 mg/dL), Rheumatoid Factors (750 IU/mL), Total Protein (21 g/dL), Albumin (60 g/L), Immunoglobulin (IgG) (60 g/L), Glucose (1000 mg/dL), Triglycerides (1584 mg/dL). A percent deviation criteria was used (not explicitly stated but implied to be within acceptable limits). |
| Cross-Reactivity | No significant cross-reactivity with specified hemoglobin fractions and glycated albumin. | Max Whole Blood Cross-Reactant Concentration with no Interference: HbA0 (120 g/dL), HbA1(a+b) (0.96 g/dL for Level 1, 1.6 g/dL for Level 2), Carbamylated Hb (2.0 g/dL), Acetylated Hb (2.0 g/dL), Glycated Albumin (10 g/dL), Labile HbA1c (1000 mg/dL). Note: Specimens with >7% HbF may yield lower than expected HbA1c values. |
| Hemoglobin Variants | Accurate results for common hemoglobin variants (HbS, HbC, HbE, HbD, HbA2) within acceptable bias. | Relative % Bias from Reference Method at Low (around 6.5%) and High (around 9%) HbA1c: - HbS: -2.5% (Low), -4.0% (High) - HbC: -3.9% (Low), -6.0% (High) - HbE: -0.1% (Low), -1.2% (High) - HbD: -1.8% (Low), -2.6% (High) - HbA2: -1.0% (Low), 0.4% (High) Note: Specimens with >7% HbF may yield lower than expected HbA1c values. |
| Exogenous Interferences (Drugs) | No significant interference from a list of commonly used drugs at specified concentrations. | No significant interference reported for 18 listed drugs (N-Acetylcysteine, Acetylsalicylic acid, Ampicillin-Na, Ascorbic acid, Cefoxitin, Heparin, Levodopa, Methyldopa + 1.5, Metronidazole, Doxycyclin, Rifampicin, Gammagard, Cyclosporine, Phenylbutazone, Acetaminophen, Ibuprofen, Theophylline, Tolbutamide) at tested concentrations. |
| Sample Matrix Comparison | Acceptable agreement across different anticoagulants and fill levels. | Mean Difference for K2-EDTA, K3-EDTA, Na Heparin, Li Heparin, NaF/Potassium oxalate, EDTA/Fluoride at full and half-full tubes indicate close agreement (e.g., -0.026 to 0.019). Upper and Lower 95% CIs are provided (e.g., -0.181 to 0.172). |
| Method Comparison | Good agreement with the NGSP reference method (Tosoh Automated Glycohemoglobin Analyzer HLC-723G8). | Whole Blood Application: Mean bias vs. NGSP TOSOH = -0.046%. Hemolysate Application: Mean bias vs. NGSP TOSOH = 0.046%. Bias at concentrations: (e.g., 5% HbA1c: WB -2.4%, Hemolysate 0.6%; 12% HbA1c: WB 0.7%, Hemolysate 1.2%). |
| Total Error (TE) | Total Error within clinical requirements, considering both bias and precision. | Hemolysate Application: TE ranging from 3.0% to 4.4% across HbA1c levels. Whole Blood Application: TE ranging from 3.1% to 5.2% across HbA1c levels. |
2. Sample size used for the test set and the data provenance
-
Precision (Repeatability and Intermediate Precision):
- Sample Size: 10 samples (2 controls, 8 human samples) for each application (Hemolysate and Whole Blood). Measured for 21 days.
- Data Provenance: Human samples and controls mentioned. No specific country of origin is stated, but implied to be laboratory-based testing. Prospective study design.
-
Analytical Sensitivity (LoB, LoD):
- Sample Size:
- LoB: One analyte-free sample. 60 measurements per lot across 3 lots.
- LoD: Five unique human samples with low-analyte concentrations. 60 measurements per lot across 3 lots.
- Data Provenance: Human samples mentioned. Laboratory-based testing. Prospective study design.
- Sample Size:
-
Linearity/Assay Reportable Range:
- Sample Size: Separate dilution series (at least eleven levels) prepared from human hemolysate sample pools.
- Data Provenance: Human hemolysate samples. Laboratory-based testing. Prospective study design.
-
Endogenous Interferences:
- Sample Size: Pooled whole blood samples at two HbA1c levels, spiked with 9 different interferents. 18 spiked samples + interferent-free pools. Each tested in ten-fold.
- Data Provenance: Pooled whole blood samples. Laboratory-based testing. Prospective study design.
-
Cross-Reactivity:
- Sample Size: Not explicitly stated as a number of unique samples, but rather a "series of experiments" with specific cross-reactants. Ten replicates of each sample were analyzed for each dilution level.
- Data Provenance: Laboratory-based testing using prepared samples to introduce cross-reactants. Prospective study design.
-
Hemoglobin Variants:
- Sample Size: 30 HbS, 30 HbC, 30 HbE, 29 HbD, 15 HbA2, 19 Elevated HbF. Total = 153 samples.
- Data Provenance: Not specified, but implied to be characterized patient samples containing the variants. Laboratory-based testing. Prospective.
-
Exogenous Interferences (Drugs):
- Sample Size: Hemolysate samples at two HbA1c levels, spiked with 18 common drugs at two concentrations. Each drug/concentration combination tested in ten-fold.
- Data Provenance: Hemolysate samples and spiked drugs. Laboratory-based testing. Prospective study design.
-
Sample Matrix Comparison:
- Sample Size: At least 40 samples of each anticoagulant type (e.g., K2-EDTA, K3-EDTA, Na Heparin, Li Heparin, NaF/Potassium oxalate, EDTA/Fluoride) at full and half-filled tubes. Samples from one donor for full/half-filled comparisons.
- Data Provenance: Human donor samples. Laboratory-based testing. Prospective study design.
-
Method Comparison:
- Sample Size: 171 whole blood samples and 173 hemolysate samples.
- Data Provenance: Samples from a "secondary NGSP reference laboratory." This suggests these are clinical samples that have been previously characterized by a gold standard (NGSP certified) method. The nature (retrospective/prospective clinical samples) is not explicitly stated but is consistent with a clinical validation using real patient samples.
3. Number of experts used to establish the ground truth for the test set and the qualifications of those experts
The document does not describe the use of human experts to establish ground truth in the way one would for image-based diagnostic AI. This device is an in vitro diagnostic (IVD) reagent system for quantifying a biomarker (HbA1c).
- Ground truth for the performance studies (e.g., precision, linearity, interferences) is established through:
- Reference materials (e.g., controls like PreciControl HbA1c norm/path).
- Dilution series from characterized sample pools.
- Spiking experiments with known concentrations of interferents or cross-reactants.
- For Hemoglobin Variants and Method Comparison:
- The "ground truth" or reference method is the NGSP Tosoh HPLC system. This system undergoes certification by the National Glycohemoglobin Standardization Program (NGSP) and is considered a highly accurate laboratory assay method, not dependent on human expert interpretation of a result. No individual human expert counts or qualifications are reported for establishing these reference values.
4. Adjudication method for the test set
Not applicable. This is an IVD device, not an AI or imaging diagnostic device that typically requires expert adjudication for ground truth establishment. The performance is assessed by comparing results to established reference methods, spiked concentrations, or statistical analysis of replicates.
5. If a multi reader multi case (MRMC) comparative effectiveness study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance
Not applicable. The Tina-quant Hemoglobin A1cDx Gen.3 assay is an in vitro diagnostic (IVD) device for quantitative determination of HbA1c. It does not involve human readers interpreting images or data with or without AI assistance. It provides a numerical result.
6. If a standalone (i.e. algorithm only without human-in-the-loop performance) was done
Yes, the studies described are all standalone (algorithm only) performance evaluations. The device autonomously measures HbA1c and Hb concentrations, calculates the ratio, and reports the final HbA1c result. There is no human intervention in the result generation process once the sample is loaded onto the analyzer.
7. The type of ground truth used
The ground truth used in various parts of the study consists of:
- Reference Methods: The NGSP Tosoh HPLC system for method comparison, which is a recognized standardized method for HbA1c determination.
- Certified Reference Materials/Controls: PreciControl HbA1c norm and path with assigned values.
- Known Concentrations: For linearity, LoB, LoD studies, the "truth" is derived from preparing samples with known concentrations or by statistical analysis of repeat measurements of low/negative samples.
- Spiked Samples: For interference and cross-reactivity studies, the "truth" is the un-spiked sample value, and the effect is measured by deviation after adding a known amount of interferent.
- Characterized Patient Samples: For hemoglobin variant testing, samples from patients known to carry specific hemoglobin variants were used.
8. The sample size for the training set
This document does not describe a "training set" in the context of machine learning or AI. The Tina-quant Hemoglobin A1cDx Gen.3 is a reagent system for a well-established immunoassay technique. Its development involved chemical and assay optimization, not machine learning model training.
9. How the ground truth for the training set was established
Not applicable, as there is no mention or indication of a machine learning "training set" for this IVD device. The assay's analytical principles are based on turbidimetric inhibition immunoassay (TINIA) and bichromatic photometric determination, which are traditional chemical analysis methods, not AI-driven algorithms requiring training data.
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March 26, 2020
Roche Diagnostics Operations Inc. Leslie Patterson Regulatory Affairs Principal 9115 Hague Rd Indianapolis, Indiana 46250
Re: K193053
Trade/Device Name: Tina-quant Hemoglobin A1cDx Gen.3 Regulation Number: 21 CFR 862.1373 Regulation Name: Hemoglobin A1c Test System Regulatory Class: Class II Product Code: PDJ, LCP Dated: February 13, 2020 Received: February 14, 2020
Dear Leslie Patterson:
We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.
If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.
Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR
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- for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safety-reportingcombination-products); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.
Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.
For comprehensive regulatory information about medical devices and radiation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).
Sincerely,
Marianela Perez-Torres, Ph.D. Acting Deputy Director Division of Chemistry and Toxicology Devices OHT7: Office of In Vitro Diagnostics and Radiological Health Office of Product Evaluation and Quality Center for Devices and Radiological Health
Enclosure
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Indications for Use
510(k) Number (if known) K193053
Device Name Tina-quant Hemoglobin A1cDx Gen.3
Indications for Use (Describe)
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 venous whole blood on the cobas c 503 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) | |
|---|---|
| ☑ Prescription Use (Part 21 CFR 801 Subpart D) | ☐ Over-The-Counter Use (21 CFR 801 Subpart C) |
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Tina-quant Hemoglobin A1cDx Gen.3 510(k) Summary (K193053)
This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of 21 CFR 807.92.
In accordance with 21 CFR 807.87, Roche Diagnostics hereby submits official notification as required by Section 510(k) of the Federal Food, Drug and Cosmetics Act of our intention to market the device described in this Premarket Notification 510(k).
The purpose of this Traditional 510(k) Premarket Notification is to obtain FDA review and clearance for the Tina-quant Hemoglobin A1cDx Gen.3 assay.
| Submitter Name | Roche Diagnostics Operations Inc. |
|---|---|
| Address | 9115 Hague RoadP.O. Box 50416Indianapolis, IN 46250-0457 |
| Contact | Leslie PattersonPhone: (317) 521-7307FAX: (317) 521-2324Email: leslie.patterson@roche.com |
| Date Prepared | February 13, 2020 |
| Proprietary Name | Tina-quant Hemoglobin A1cDx Gen.3 |
| Common Name | Glycosylated Hemoglobin Assay |
| Classification Name | Hemoglobin A1c test system |
| Product Codes,Regulation Numbers | PDJ, 862.1373LCP, 864.7470 |
| Predicate Device | COBAS INTEGRA 800 Tina-quant Hemoglobin A1cDx Gen.2 assay, K121291 |
| Establishment Registration | 1823260, Roche Diagnostics Corporation |
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1. DEVICE DESCRIPTION
Tina-quant Hemoglobin A1cDx Gen.3 assay 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 cobas c 503 clinical chemistry analyzer.
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 and hemolysis occurs onboard the analyzer. For the Hemolysate Application, hemolyzed samples are placed on the analyzer and hemolysis occurs manually before placing the samples onboard the analyzer. The two applications yield the same results. 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.
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.
Test Principle 1.1.
This method uses tetradecyltrimethylammonium bromide (TTAB) as the detergent in the hemolyzing reagent to eliminate interference from leukocytes (TTAB does not lyse leukocytes). Sample pretreatment to remove labile HbA1c is not necessary. All hemoglobin variants which are glycated at the ß -chain 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 (HbAC, HbAC, HbAD) can be determined using this assay.
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1.2. Hemoglobin A1c
The HbA1c determination is based on the turbidimetric inhibition immunoassay (TINIA) for hemolyzed whole blood.
-
. 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 R3 (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.
1.3. 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.
Final HbA1c Result 1.4.
The final result is expressed as mmol/mol 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
1.5. 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.
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2. INDICATIONS FOR USE
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 venous whole blood on the cobas c 503 clinical chemistry analyzer. HbA1c determinations are useful for monitoring of long-term blood glucose control in individuals with diabetes mellitus.
TECHNOLOGICAL CHARACTERISTICS 3.
The following table compares the Tina-quant Hemoglobin A1cDx Gen.3 assay with its predicate device, COBAS INTEGRA 800 Tina-quant Hemoglobin A1cDx Gen.2 (K121291).
| Feature | Predicate Device:COBAS INTEGRA 800 Tina-quantHemoglobin A1cDx Gen.2 K121291 | Submitted Device:Tina-quant Hemoglobin A1cDx Gen.3 |
|---|---|---|
| Intended Use | This test is to be used as an aid in diagnosis ofdiabetes and as an aid in identifying patientswho may be at risk for developing diabetes. Itis an in vitro diagnostic reagent systemintended for quantitative determination ofmmol/mol hemoglobin A1c (IFCC) and %hemoglobin A1c (DCCT/NGSP) in hemolysateor venous whole blood. HbA1c determinationsare useful for monitoring of long-term bloodglucose control in individuals with diabetesmellitus. | Same |
| Sample Types | Anticoagulated venous bloodAcceptable anticoagulants for both thehemolysate and whole blood applicationsinclude:• Li-Heparin• K2-EDTA• K3-EDTA• Fluoride/potassium oxalate• Na-Heparin• NaF/Na2-EDTA | Anticoagulated venous bloodAcceptable anticoagulants for both thehemolysate and whole blood applicationsinclude:• Li-Heparin• K2-EDTA• K3-EDTA• Fluoride/potassium oxalate• Na-Heparin• EDTA Fluoride |
| InstrumentPlatform | COBAS Integra 800• Absorbance Photometry | cobas c 503 |
| Calibrator | Cfas HbA1c | Same |
| Feature | Predicate Device:COBAS INTEGRA 800 Tina-quantHemoglobin A1cDx Gen.2 K121291 | Submitted Device:Tina-quant Hemoglobin A1cDx Gen.3 |
| CalibrationFrequency | • After 29 days on-board the analyzer• After reagent lot change• As required following quality controlprocedures | Same |
| Calibration Mode | Logit/Log 5 | Hb: linearHbA1c : RCM4 |
| Controls | PreciControl HbA1c normPreciControl HbA1c path | Same |
| Reagent Stability | Unopened:• 2-8 °C until expiration dateOn-board in use:• 2-8 °C for 28 days | Same |
| Reporting Units | • % HbA1c (NGSP/DCCT)• mmol/mol (IFCC) | Same |
| Antibody | Polyclonal anti-HbA1c from sheep blood | Same |
| Test Principle | The anticoagulated whole blood specimen ishemolyzed automatically on the COBASINTEGRA 800 analyzers with COBASINTEGRA Hemolyzing Reagent Gen.2. Thismethod uses TTAB as the detergent in thehemolyzing reagent to eliminate interferencefrom leukocytes (TTAB does not lyseleukocytes). Sample pretreatment to removelabile HbA1c is not necessary. All hemoglobinvariants which are glycated at the β-chain N-terminus and which have antibody-recognizableregions identical to that of HbA1c aredetermined by this assay. Consequently, themetabolic state of diabetic patients havinguremia or the most frequenthemoglobinopathies (HbAS, HbAC, HbAE,HbAD) can be determined by this assay. | This method usestetradecyltrimethylammonium bromide (TTAB)as the detergent in the hemolyzing reagent toeliminate interference from leukocytes (TTABdoes not lyse leukocytes). Samplepretreatment to remove labile HbA1c is notnecessary. All hemoglobin variants which areglycated at the β -chain N-terminus and whichhave antibody-recognizable regions identical tothat of HbA1c are determined by this assay.Consequently, the metabolic state of patientshaving uremia or the most frequenthemoglobinopathies (HbAS, HbAC, HbAE,HbAD) can be determined using this assay. |
| Determination ofHbA1c | Turbidimetric immunoinhibition (TINIA).Antigen-antibody complexes are formedand excess Ab aggregate with polyhapten toform insoluble complexes | HbA1c determination is based on theturbidimetric inhibition immunoassay (TINIA)for hemolyzed whole blood. Glycohemoglobinin the sample reacts with anti-HbA1c antibodyto form soluble antigen-antibody complexes.Polyhaptens react with excess anti-HbA1cantibodies to form an insoluble antibody-polyhapten complex which can be measuredturbidimetrically. |
| Determination ofHb | Bichromatic photometric determinationafter conversion to a colored derivate | Liberated hemoglobin in the hemolyzedsample is converted to a derivative having acharacteristic absorption spectrum which ismeasured bichromatically. |
| Feature | Predicate Device:COBAS INTEGRA 800 Tina-quantHemoglobin A1cDx Gen.2 K121291 | Submitted Device:Tina-quant Hemoglobin A1cDx Gen.3 |
| Determinationof % HbA1c | The final result is expressed as % HbA1c andis calculated from the HbA1c/Hb ratio perDCCT/NGSP as follows:$HbA1c (%) = (HbA1c/Hb) × 91.5 + 2.15$ | Same |
| Measuring Range | Hemoglobin: 4-35 g/dLHbA1c: 0.3-3.4 g/dLThis corresponds to a measuring range of23-258 mmol/mol HbA1c (IFCC)and 4.3-24.8 % HbA1c (DCCT/NGSP) at atypical hemoglobin concentration of 13.2 g/dL. | Hemoglobin: 4-40 g/dL (2.48-24.8 mmol/L)HbA1c: 0.3-2.6 g/dL (0.186-1.61 mmol/L)This corresponds to a measuring range of 23-196 mmol/molHbA1c (IFCC) and 4.2-20.1 %HbA1c (DCCT/NGSP) at a typical hemoglobinconcentration of 13.2 g/dL (8.2 mmol/L). |
| Traceability | The assigned HbA1c and total hemoglobinvalues of the cobas c Tina-quant HemoglobinA1cDx Gen.3 assay is certified with theNational Glycohemoglobin StandardizationProgram (NGSP). NGSP certification isrepeated annually. | Same |
| ReagentComposition | 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; preservativesSR Polyhapten Reagent:• MES buffer: 0.025 mol/L• TRIS buffer: 0.015 mol/L, pH 6.2• HbA1c polyhapten: > 8µg/mL• stabilizers; preservativesA1CD (Hemolyzing Reagent):• Aqueous buffered matrix, pH 7.25• Tetradecyltrimethylammonium bromide: 36g/L• sodium dihydrogenphosphate monohydrate:16 mmol/L• sodium monohydrogenphosphate dihydrate:64 mmol/Lstabilizers; preservatives | R1 Antibody Reagent:•MES (2-morpholinoethane sulfonicacid) 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; preservativesR3 Polyhapten Reagent:•MES buffer: 0.025 mol/L•TRIS buffer: 0.015 mol/L, pH 6.2•HbA1c polyhapten: > 8 µg/mL• detergents; stabilizers; preservativesA1CD (Hemolyzing Reagent):• Aqueous buffered matrix, pH 7.25• Tetradecyltrimethylammonium bromide: 36g/L• Sodium dihydrogenphosphate monohydrate:16 mmol/L• Sodium monohydrogenphosphate dihydrate:64 mmol/Lstabilizers; preservatives |
Table 1: Substantial Equivalence Assay Comparison
{7}------------------------------------------------
{8}------------------------------------------------
{9}------------------------------------------------
4. NON-CLINICAL PERFORMANCE EVALUATION
Performance characteristics were evaluated with Tina-quant Hemoglobin A1cDx Gen.3 on the cobas c 503 analytical unit.
Tina-quant Hemoglobin A1cDx Gen.3 offers two sample type specific applications, Hemolysate Application and Whole Blood Application, one for manually hemolyzed samples and one for whole blood samples respectively.
The Tina-quant Hemoglobin A1cDx Gen.3 assay first measures total hemoglobin (Hb) and glycated hemoglobin (HbA1c) in terms of either g/dL or mmol/L. Then the analyzer calculates the HbA1c/Hb ratio according to 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 included limit of detection and linearity. These results were reported in terms of Hb and HbA1c individually. Patient sample values are reported in terms of the ratio of glycated to total hemoglobin. Method comparison, control recovery and precision were evaluated in terms of the ratio.
The following performance data were provided in support of the substantial equivalence determination:
Precision 4.1.
Repeatability and Intermediate Precision 4.1.1.
Precision measurements were conducted to evaluate repeatability (within-run precision) and intermediate precision (within-laboratory precision) according the CLSI guideline EP5-A3.
Two aliquots per sample were measured once each, in two runs per day, for 21 days, on 3 cobas c 503 analytical units and using 3 reagent lots per system. Ten total samples were evaluated in each run: two controls, PreciControl HbA1c norm and PreciControl HbA1c path, and eight human samples with approximate Hb1Ac concentrations of 4.9%, 6.6%, 7.3%, 8.2%, 12.5%, 14.6%, 12.3% and 13.1% for Whole Blood and 5.0%, 6.6%, 7.3%, 8.3%, 12.5%, 14.7%, 12.1% and 12.9% for Hemolysate.
{10}------------------------------------------------
The samples were randomized within each run. 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.
| Mean% HbA1c | Repeatability (error) | Between-Run | Between-Day | Between-lot | Intermediate Precision(total) | |||||
|---|---|---|---|---|---|---|---|---|---|---|
| SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | |
| Hem 14.96 | 0.025 | 0.5 | 0.006 | 0.1 | 0.071 | 1.4 | 0.016 | 0.3 | 0.077 | 1.6 |
| Hem 26.62 | 0.027 | 0.4 | 0.013 | 0.2 | 0.053 | 0.8 | 0.059 | 0.9 | 0.085 | 1.3 |
| Hem 37.32 | 0.035 | 0.5 | 0.000 | 0.0 | 0.053 | 0.7 | 0.067 | 0.9 | 0.092 | 1.3 |
| Hem 48.32 | 0.039 | 0.5 | 0.009 | 0.1 | 0.056 | 0.7 | 0.083 | 1.0 | 0.108 | 1.3 |
| Hem 512.54 | 0.057 | 0.5 | 0.011 | 0.1 | 0.100 | 0.8 | 0.203 | 1.6 | 0.234 | 1.9 |
| HE 00614.77 | 0.077 | 0.5 | 0.013 | 0.1 | 0.148 | 1.0 | 0.268 | 1.8 | 0.316 | 2.1 |
| HE_00712.14 | 0.055 | 0.5 | 0.023 | 0.2 | 0.100 | 0.8 | 0.181 | 1.5 | 0.215 | 1.8 |
| HE_00812.94 | 0.072 | 0.6 | 0.000 | 0.0 | 0.111 | 0.9 | 0.188 | 1.5 | 0.230 | 1.8 |
| PCA1N5.53 | 0.024 | 0.4 | 0.009 | 0.2 | 0.059 | 1.1 | 0.028 | 0.5 | 0.071 | 1.3 |
| PCA1P10.89 | 0.055 | 0.5 | 0.026 | 0.2 | 0.085 | 0.8 | 0.146 | 1.3 | 0.179 | 1.6 |
Table 2: Precision Results – Hemolysate Application, Analytical Unit 1
| Table 3: Precision Results - Hemolysate Application, Analytical Unit 2 | |||
|---|---|---|---|
| Mean% HbA1c | Repeatability (error) | Between-Run | Between-Day | Between-lot | Intermediate Precision(total) | |||||
|---|---|---|---|---|---|---|---|---|---|---|
| SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | |
| Hem 14.96 | 0.027 | 0.5 | 0.005 | 0.1 | 0.034 | 0.7 | 0.015 | 0.3 | 0.046 | 0.9 |
| Hem 26.59 | 0.035 | 0.5 | 0.000 | 0.0 | 0.038 | 0.6 | 0.057 | 0.9 | 0.077 | 1.2 |
| Hem 37.29 | 0.041 | 0.6 | 0.000 | 0.0 | 0.043 | 0.6 | 0.068 | 0.9 | 0.090 | 1.2 |
| Hem 48.28 | 0.039 | 0.5 | 0.015 | 0.2 | 0.046 | 0.6 | 0.093 | 1.1 | 0.112 | 1.4 |
| Hem 512.43 | 0.069 | 0.6 | 0.027 | 0.2 | 0.038 | 0.3 | 0.175 | 1.4 | 0.193 | 1.6 |
| HE 00614.68 | 0.085 | 0.6 | 0.011 | 0.1 | 0.060 | 0.4 | 0.220 | 1.5 | 0.243 | 1.7 |
| HE_00712.05 | 0.063 | 0.5 | 0.018 | 0.1 | 0.036 | 0.3 | 0.163 | 1.4 | 0.179 | 1.5 |
| HE_00812.85 | 0.071 | 0.6 | 0.034 | 0.3 | 0.053 | 0.4 | 0.177 | 1.4 | 0.201 | 1.6 |
| PCA1N5.52 | 0.030 | 0.5 | 0.008 | 0.1 | 0.029 | 0.5 | 0.024 | 0.4 | 0.049 | 0.9 |
| PCA1P10.81 | 0.074 | 0.7 | 0.000 | 0.0 | 0.041 | 0.4 | 0.134 | 1.2 | 0.159 | 1.5 |
Table 4: Precision Results – Hemolysate Application, Analytical Unit 3
| Mean %HbA1c | Repeatability (error)SD | Repeatability (error)CV (%) | Between-RunSD | Between-RunCV (%) | Between-DaySD | Between-DayCV (%) | Between-lotSD | Between-lotCV (%) | Intermediate Precision (total)SD | Intermediate Precision (total)CV (%) |
|---|---|---|---|---|---|---|---|---|---|---|
| Hem 14.94 | 0.027 | 0.5 | 0.010 | 0.2 | 0.031 | 0.6 | 0.023 | 0.5 | 0.048 | 1.0 |
| Hem 26.57 | 0.030 | 0.5 | 0.004 | 0.1 | 0.035 | 0.5 | 0.049 | 0.7 | 0.067 | 1.0 |
{11}------------------------------------------------
| Hem 37.28 | 0.032 | 0.4 | 0.000 | 0.0 | 0.038 | 0.5 | 0.054 | 0.7 | 0.073 | 1.0 |
|---|---|---|---|---|---|---|---|---|---|---|
| Hem 48.28 | 0.040 | 0.5 | 0.000 | 0.0 | 0.041 | 0.5 | 0.070 | 0.8 | 0.091 | 1.1 |
| Hem 512.43 | 0.063 | 0.5 | 0.024 | 0.2 | 0.045 | 0.4 | 0.162 | 1.3 | 0.181 | 1.5 |
| HE 00614.68 | 0.075 | 0.5 | 0.000 | 0.0 | 0.050 | 0.3 | 0.240 | 1.6 | 0.256 | 1.7 |
| HE 00712.07 | 0.064 | 0.5 | 0.000 | 0.0 | 0.039 | 0.3 | 0.146 | 1.2 | 0.164 | 1.4 |
| HE_00812.84 | 0.072 | 0.6 | 0.000 | 0.0 | 0.052 | 0.4 | 0.150 | 1.2 | 0.174 | 1.4 |
| PCA1N5.49 | 0.026 | 0.5 | 0.008 | 0.2 | 0.035 | 0.6 | 0.024 | 0.4 | 0.050 | 0.9 |
| PCA1P10.78 | 0.071 | 0.7 | 0.000 | 0.0 | 0.048 | 0.4 | 0.116 | 1.1 | 0.144 | 1.3 |
| Table 5: Precision Results - Hemolysate Application, 3 Lots and 3 Analytical Units | |
|---|---|
| ------------------------------------------------------------------------------------ | -- |
| Mean,%HbA1c | Repeatability(error) | Between-Run | Between-Day | Between-Lot | Between-Device | Reproducibility(total) | |||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| SD | CV(%) | SD | CV(%) | SD | CV(%) | SD | CV(%) | SD | CV(%) | SD | CV(%) | ||
| Hem 1,4.96 | 0.026 | 0.5 | 0.007 | 0.1 | 0.049 | 1.0 | 0.019 | 0.4 | 0.009 | 0.2 | 0.059 | 1.2 | |
| Hem 2,6.59 | 0.031 | 0.5 | 0.006 | 0.1 | 0.042 | 0.6 | 0.055 | 0.8 | 0.023 | 0.3 | 0.080 | 1.2 | |
| Hem 3,7.30 | 0.036 | 0.5 | 0.000 | 0.0 | 0.045 | 0.6 | 0.063 | 0.9 | 0.019 | 0.3 | 0.088 | 1.2 | |
| Hem 4,8.29 | 0.039 | 0.5 | 0.005 | 0.1 | 0.049 | 0.6 | 0.082 | 1.0 | 0.019 | 0.2 | 0.105 | 1.3 | |
| Hem 5,12.47 | 0.063 | 0.5 | 0.022 | 0.2 | 0.070 | 0.6 | 0.179 | 1.4 | 0.061 | 0.5 | 0.212 | 1.7 | |
| HE_006,14.71 | 0.079 | 0.5 | 0.010 | 0.1 | 0.098 | 0.7 | 0.242 | 1.6 | 0.053 | 0.4 | 0.278 | 1.9 | |
| HE_007,12.08 | 0.061 | 0.5 | 0.016 | 0.1 | 0.067 | 0.6 | 0.163 | 1.3 | 0.048 | 0.4 | 0.193 | 1.6 | |
| HE_008,12.88 | 0.072 | 0.6 | 0.017 | 0.1 | 0.078 | 0.6 | 0.172 | 1.3 | 0.055 | 0.4 | 0.210 | 1.6 | |
| PCA1N,5.51 | 0.027 | 0.5 | 0.008 | 0.2 | 0.043 | 0.8 | 0.027 | 0.5 | 0.022 | 0.4 | 0.062 | 1.1 | |
| PCA1P,10.83 | 0.067 | 0.6 | 0.000 | 0.0 | 0.062 | 0.6 | 0.132 | 1.2 | 0.054 | 0.5 | 0.169 | 1.6 |
Table 6: Precision Results – Whole Blood Application, Analytical Unit 1
| Mean | Repeatability (error) | Between-Run | Between-Day | Between-Lot | Intermediate Precision(total) | |||||
|---|---|---|---|---|---|---|---|---|---|---|
| %HbA1c | SD | CV(%) | SD | CV(%) | SD | CV(%) | SD | CV(%) | SD | CV(%) |
| WB 1,4.87 | 0.038 | 0.8 | 0.000 | 0.0 | 0.043 | 0.9 | 0.045 | 0.9 | 0.073 | 1.5 |
| WB 2,6.60 | 0.026 | 0.4 | 0.015 | 0.2 | 0.030 | 0.4 | 0.071 | 1.1 | 0.083 | 1.3 |
| WB 3,7.37 | 0.032 | 0.4 | 0.018 | 0.2 | 0.032 | 0.4 | 0.084 | 1.1 | 0.097 | 1.3 |
| WB 4,8.24 | 0.039 | 0.5 | 0.008 | 0.1 | 0.048 | 0.6 | 0.089 | 1.1 | 0.108 | 1.3 |
| WB 5,12.59 | 0.056 | 0.4 | 0.025 | 0.2 | 0.061 | 0.5 | 0.154 | 1.2 | 0.177 | 1.4 |
| WB_006,14.69 | 0.079 | 0.5 | 0.043 | 0.3 | 0.067 | 0.5 | 0.168 | 1.1 | 0.202 | 1.4 |
| WB_007,12.34 | 0.062 | 0.5 | 0.032 | 0.3 | 0.057 | 0.5 | 0.152 | 1.2 | 0.176 | 1.4 |
| WB_008,13.14 | 0.061 | 0.5 | 0.015 | 0.1 | 0.055 | 0.4 | 0.165 | 1.3 | 0.185 | 1.4 |
| PCA1N,5.51 | 0.029 | 0.5 | 0.002 | 0.0 | 0.039 | 0.7 | 0.044 | 0.8 | 0.066 | 1.2 |
| PCA1P,11.20 | 0.055 | 0.5 | 0.011 | 0.1 | 0.057 | 0.5 | 0.129 | 1.2 | 0.152 | 1.4 |
{12}------------------------------------------------
| Mean%HbA1c | Repeatability (error) | Between-Run | Between-Day | Between-Lot | Intermediate Precision(total) | |||||
|---|---|---|---|---|---|---|---|---|---|---|
| SD | CV(%) | SD | CV(%) | SD | CV(%) | SD | CV(%) | SD | CV(%) | |
| WB 1,4.88 | 0.032 | 0.7 | 0.013 | 0.3 | 0.026 | 0.5 | 0.031 | 0.6 | 0.053 | 1.1 |
| WB 2,6.58 | 0.031 | 0.5 | 0.010 | 0.2 | 0.025 | 0.4 | 0.077 | 1.2 | 0.087 | 1.3 |
| WB 3,7.35 | 0.040 | 0.5 | 0.000 | 0.0 | 0.029 | 0.4 | 0.091 | 1.2 | 0.103 | 1.4 |
| WB 4,8.21 | 0.042 | 0.5 | 0.011 | 0.1 | 0.035 | 0.4 | 0.106 | 1.3 | 0.120 | 1.5 |
| WB 5,12.53 | 0.070 | 0.6 | 0.029 | 0.2 | 0.047 | 0.4 | 0.164 | 1.3 | 0.186 | 1.5 |
| WB_006,14.62 | 0.095 | 0.6 | 0.035 | 0.2 | 0.091 | 0.6 | 0.198 | 1.4 | 0.240 | 1.6 |
| WB_007,12.25 | 0.078 | 0.6 | 0.000 | 0.0 | 0.035 | 0.3 | 0.176 | 1.4 | 0.195 | 1.6 |
| WB_008,13.06 | 0.073 | 0.6 | 0.009 | 0.1 | 0.052 | 0.4 | 0.163 | 1.2 | 0.186 | 1.4 |
| PCA1N,5.51 | 0.036 | 0.6 | 0.000 | 0.0 | 0.028 | 0.5 | 0.044 | 0.8 | 0.063 | 1.1 |
| PCA1P,11.13 | 0.062 | 0.6 | 0.033 | 0.3 | 0.043 | 0.4 | 0.162 | 1.5 | 0.182 | 1.6 |
Table 7: Precision Results – Whole Blood Application, Analytical Unit 2
| Table 8: Precision Results - Whole Blood Application, Analytical Unit 3 | |||||||
|---|---|---|---|---|---|---|---|
| -- | -- | -- | -- | -- | -------------------------------------------------------------------------- | -- | -- |
| Mean | Repeatability (error) | Between-Run | Between-Day | Between-Lot | Intermediate Precision(total) | |||||
|---|---|---|---|---|---|---|---|---|---|---|
| %HbA1c | SD | CV(%) | SD | CV(%) | SD | CV(%) | SD | CV(%) | SD | CV(%) |
| WB 1,4.86 | 0.029 | 0.6 | 0 | 0.0 | 0.035 | 0.7 | 0.044 | 0.9 | 0.064 | 1.3 |
| WB 2,6.55 | 0.031 | 0.5 | 0.009 | 0.1 | 0.025 | 0.4 | 0.080 | 1.2 | 0.090 | 1.4 |
| WB 3,7.31 | 0.039 | 0.5 | 0.006 | 0.1 | 0.026 | 0.4 | 0.090 | 1.2 | 0.101 | 1.4 |
| WB 4,8.17 | 0.043 | 0.5 | 0.012 | 0.2 | 0.036 | 0.4 | 0.096 | 1.2 | 0.111 | 1.4 |
| WB 5,12.51 | 0.079 | 0.6 | 0.000 | 0.0 | 0.064 | 0.5 | 0.154 | 1.2 | 0.184 | 1.5 |
| WB_006,14.62 | 0.082 | 0.6 | 0.000 | 0.0 | 0.107 | 0.7 | 0.191 | 1.3 | 0.234 | 1.6 |
| WB_007,12.23 | 0.068 | 0.6 | 0.028 | 0.2 | 0.056 | 0.5 | 0.157 | 1.3 | 0.182 | 1.5 |
| WB_008,13.05 | 0.083 | 0.6 | 0.000 | 0.0 | 0.064 | 0.5 | 0.162 | 1.2 | 0.193 | 1.5 |
| PCA1N,5.50 | 0.030 | 0.6 | 0.006 | 0.1 | 0.028 | 0.5 | 0.051 | 0.9 | 0.066 | 1.2 |
| PCA1P,11.10 | 0.071 | 0.6 | 0.000 | 0.0 | 0.043 | 0.4 | 0.137 | 1.2 | 0.160 | 1.4 |
{13}------------------------------------------------
| Mean | Repeatability(error) | Between-Run | Between-Day | Between-Lot | Between-Device | Reproducibility(total) | |||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| %HbA1c | SD | CV(%) | SD | CV(%) | SD | CV(%) | SD | CV(%) | SD | CV(%) | SD | CV(%) | |
| WB 1,4.87 | 0.034 | 0.7 | 0.004 | 0.1 | 0.036 | 0.7 | 0.040 | 0.8 | 0.007 | 0.1 | 0.064 | 1.3 | |
| WB 2,6.57 | 0.029 | 0.4 | 0.012 | 0.2 | 0.028 | 0.4 | 0.075 | 1.1 | 0.024 | 0.4 | 0.090 | 1.4 | |
| WB 3,7.34 | 0.037 | 0.5 | 0.009 | 0.1 | 0.032 | 0.4 | 0.087 | 1.2 | 0.027 | 0.4 | 0.104 | 1.4 | |
| WB 4,8.20 | 0.041 | 0.5 | 0.011 | 0.1 | 0.042 | 0.5 | 0.096 | 1.2 | 0.031 | 0.4 | 0.117 | 1.4 | |
| WB 5,12.54 | 0.069 | 0.5 | 0.020 | 0.2 | 0.062 | 0.5 | 0.155 | 1.2 | 0.043 | 0.3 | 0.187 | 1.5 | |
| WB_006,14.64 | 0.086 | 0.6 | 0.028 | 0.2 | 0.096 | 0.7 | 0.181 | 1.2 | 0.043 | 0.3 | 0.228 | 1.6 | |
| WB_007,12.27 | 0.070 | 0.6 | 0.023 | 0.2 | 0.055 | 0.5 | 0.159 | 1.3 | 0.055 | 0.4 | 0.192 | 1.6 | |
| WB_008,13.08 | 0.073 | 0.6 | 0.000 | 0.0 | 0.061 | 0.5 | 0.161 | 1.2 | 0.049 | 0.4 | 0.194 | 1.5 | |
| PCA1N,5.51 | 0.032 | 0.6 | 0.002 | 0.0 | 0.032 | 0.6 | 0.047 | 0.8 | 0.007 | 0.1 | 0.065 | 1.2 | |
| PCA1P,11.14 | 0.063 | 0.6 | 0.019 | 0.2 | 0.054 | 0.5 | 0.141 | 1.3 | 0.047 | 0.4 | 0.171 | 1.5 |
Table 9: Precision Results – Whole Blood Application, 3 Lots and 3 Analytical Units
Analytical Sensitivity 4.2.
Limit of Blank (LoB) 4.2.1.
For determination of LoB, one analyte free sample was measured with three lots of Tina-quant Hemoglobin A1cDx Gen.3, in 10-fold determinations. Six runs were distributed over ≥ three days and measured using one cobas c 503 analytical unit. In total, 60 measurements were obtained per lot. Data analysis was 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.
LoB was determined according to CLSI guideline EP17-A2.
Table 10: LoB Results
| Hb LoB | HbA1c LoB | |||
|---|---|---|---|---|
| mmol/L | g/dL | mmol/L | g/dL | |
| 0.0530 mmol/L | 0.085 g/dL | 0.0220 mmol/L | 0.035 g/dL |
This corresponds to a Limit of Blank of 15 mmol/mol (IFCC) and 3.5 % HbA1c (DCCT/NGSP) at a typical hemoglobin concentration of 13.2 g/dL (8.2 mmol/L).
{14}------------------------------------------------
Limit of Detection (LoD) 4.2.2.
For determination of LoD, five unique human samples with low-analyte concentrations were measured with three lots of Tina-quant Hemoglobin A1cDx Gen.3 in two-fold determinations. The measurements were performed in six runs, over > three days, on one cobas c 503 analytical unit. In total. 60 measurements were obtained per lot.
LoD is defined as the concentration, at which there is a 95% probability that a sample contains analyte.
LoD was determined according to CLSI guideline EP17-A2.
Table 11: LoD Results
| Hb LoD | HbA1c LoD | ||
|---|---|---|---|
| mmol/L | g/dL | mmol/L | g/dL |
| 0.119 mmol/L | 0.192 g/dL | 0.0437 mmol/L | 0.07 g/dL |
This corresponds to a Limit of Detection of 22 mmol/mol (IFCC) and 4.2 % HbA1c (DCCT/NGSP) at a typical hemoglobin concentration of 13.2 g/dL (8.2 mmol/L).
Linearity/Assay Reportable Range 4.3.
Separate dilution series, consisting of at least eleven levels, were prepared for each glycated hemoglobin (HbA1c) and total hemoglobin (Hb) using human hemolysate sample pools. The sample pools include HbA1c and Hb 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 performed separately for each sample.
The study was performed according to CLSI guideline EP6-A.
Table 12: Linearity Results according to CLSI EP6-A
| Application | Analyte | Low End of Linear Range | High End of Linear Range | ||
|---|---|---|---|---|---|
| g/dL | mmol/L | g/dL | mmol/L | ||
| Hemolysate | Hb | 3.04 | 1.89 | 40.4 | 25.1 |
| HbA1c | 0.293 | 0.182 | 2.87 | 1.78 |
{15}------------------------------------------------
Table 13: Empirical First Order Regression Results
| Application | Analyte | Slope | Intercept | Pearson's r |
|---|---|---|---|---|
| Hemolysate | Hb | 1.019 | -0.1552 | 0.9999 |
| HbA1c | 0.991 | -0.0026 | 0.9990 |
Linearity was determined throughout the claimed measuring range of:
4 – 40 g/dL (2.48 – 24.8 mmol/L) Hb:
0.3 – 2.6 g/dL (0.186 – 1.61 mmol/L) HbA1c:
This corresponds to a measuring range of 23-196 mmol/mol HbA1c (IFCC) and 4.2-20.1 % HbA1c (DCCT/NGSP) at a typical hemoglobin concentration of 13.2 g/dL (8.2 mmol/L).
4.4. Endogenous Interferences
A study evaluated several endogenous substances for potential interference with measurement of % HbA1c. The following nine endogenous substances were evaluated.
- Bilirubin .
- Ditaurobilirubin .
- Lipemia .
- Rheumatoid Factors .
- Total Protein .
- Albumin .
- Immunoglubulin (IgG) .
- Glucose .
- Triglycerides .
Pooled whole blood samples, with two hemoglobin A1c levels, one near the medical decision level and one above it, were spiked with the maximum level of the above nine interferents, in separate preparations, resulting in eighteen spiked samples were then hemolyzed
{16}------------------------------------------------
with Tina-quant HbA1c Hemolyzing Reagent. Another pool, without interferent, was equally hemolyzed. A > ten-level dilution series was created for each of the eighteen spiked samples, using the interferent-free pools as the diluent.
The eighteen dilution series were tested in ten-fold, using one reagent lot, one cobas c 503 analytical unit, in a single run and within one calibration cycle. Additionally, PreciControl HbA1c norm and PreciControl HbA1c path were used as the controls. The mean of the ten replicates was compared to the result from the reference sample (aliquot with no interferent). The comparison was evaluated as a percent deviation. For purposes of this experiment, the data was collected using the Hemolysate Application and was representative of both Hemolysate and Whole Blood Applications.
| Potential Interferent | Claimed Maximum Concentration without Interference |
|---|---|
| Bilirubin | 60 mg/dL |
| Ditaurobilirubin | 60 mg/dL |
| Lipemia | 400 mg/dL |
| Rheumatoid Factors | 750 IU/mL |
| Total Protein | 21 g/dL |
| Albumin | 60 g/L |
| Immunoglobulin (IgG) | 60 g/L |
| Glucose | 1000 mg/dL |
| Triglycerides | 1584 mg/dL |
Table 14: Endogenous Interference
4.5. Cross-Reactivity
This study was conducted to evaluate the Tina-quant Hemoglobin A1cDx Gen.3 assay on the cobas c 503 analytical unit for potential cross-reactivity with the following hemoglobin fractions and glycated albumin.
- HbA0 .
- Carbamylated Hb .
- HbA1(a+b) .
- Acetylated Hb .
- Labile HbA1c Glycated Albumin . .
{17}------------------------------------------------
A series of experiments were performed using one reagent lot, on one cobas c 503 analytical unit, in a single run, within one calibration cycle. PreciControl HbA1c norm and PreciControl HbA1c path were used as the controls. Ten replicates of each sample were analyzed for each dilution level. The median % HbA1c of each dilution level was compared to the median % HbA1c from dilution level zero (without cross-reactant). For purposes of this experiment, the data was collected using the Hemolysate Application and was representative of both Hemolysate and Whole Blood Applications.
| Cross-Reactant | Max Whole Blood Cross-Reactant Concentration | Max Whole Blood Cross-Reactant Concentration withno Interference | |
|---|---|---|---|
| HbA0 | HbA1c Level 1 | 120 g/dL | 120 g/dL |
| HbA1c Level 2 | 120 g/dL | 120 g/dL | |
| HbA1(a+b) | HbA1c Level 1 | 1.6 g/dL | 0.96 g/dL |
| HbA1c Level 2 | 1.6 g/dL | 1.6 g/dL | |
| Carbamylated Hb | HbA1c Level 1 | 2.0 g/dL | 2.0 g/dL |
| HbA1c Level 2 | 2.0 g/dL | 2.0 g/dL | |
| Acetylated Hb | HbA1c Level 1 | 2.0 g/dL | 2.0 g/dL |
| HbA1c Level 2 | 2.0 g/dL | 2.0 g/dL | |
| Glycated Albumin | HbA1c Level 1 | 10 g/dL | 10 g/dL |
| HbA1c Level 2 | 10 g/dL | 10 g/dL | |
| Labile HbA1c | HbA1c Level 1 | 1000 mg/dL | 1000 mg/dL |
| HbA1c Level 2 | 1000 mg/dL | 1000 mg/dL |
Table 15: Cross-Reactivity
Hemoglobin Variants 4.6.
Hemoglobin variant testing was conducted to determine if significant interference with any of the major hemoglobin variants occurred when using the Tina-quant Hemoglobin A1cDx Gen.3 assay on cobas c 503 analytical unit. 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. Additionally, in some conditions fetal hemoglobin, HbF, is elevated. Also, the erythrocytes of some patients (e.g. beta thalassemia minor) contain elevated levels of HbA2.
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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 | 30 | 35-41% S | 4.35 - 12.7 |
| HbC | 30 | 28-37% C | 4.90 - 14.1 |
| HbE | 30 | 24-27% E | 5.17 - 10.0 |
| HbD | 29 | 36-42% D | 5.17 - 9.70 |
| HbA2 | 15 | 4.3-6.5% A2 | 5.10 - 9.80 |
| Elevated HbF | 19 | 3.2-39% F | 6.10 - 9.30 |
Table 16: Hemoglobin Variant Samples
Each sample was tested once, in at least one run, on one cobas c 503 analytical unit. Results obtained with the Tina-quant Hemoglobin A1cDx Gen.3 assay on the cobas c 503 analytical unit were compared to those obtained with the corresponding reference method. For purposes of this experiment, the data was collected using the Hemolysate Application and was representative of both Hemolysate and Whole Blood Applications.
Table 17: Hemoglobin Variant Testing
| Percent Relative Bias from Reference Method at Low and High Concentrations of HbA1c Samples | ||||
|---|---|---|---|---|
| Around 6.5% HbA1c | Around 9% HbA1c | |||
| HbVariant | Relative % Difference | Range | Relative % Difference | Range |
| HbS | -2.5 | -7.2 - 3.2 | -4.0 | -9.3 - (-2.0) |
| HbC | -3.9 | -7.7 - 2.8 | -6.0 | -4.6 - (-3.6) |
| HbE | -0.1 | -5.5 - 5.7 | -1.2 | -5.2 - 0.6 |
| HbD | -1.8 | -4.5 - 3.0 | -2.6 | -3.3 - 0.2 |
| HbA2 | -1.0 | -4.1 - 2.7 | 0.4 | -2.2 - 1.1 |
| HbF | Specimens containing high amounts of HbF (>7%) may yield lower than expected HbA1c values. |
Exogenous Interferences – Drugs 4.7.
The purpose of this study was to evaluate drugs for potential interference with the Tina-quant Hemoglobin A1cDx Gen.3 assay measured on the cobas c 503 analytical unit.
The eighteen commonly used drugs listed below were added to samples and examined for potential effect on % HbA1c determination. Drug interference testing was performed with
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hemolysate samples at two different HbA1c levels, approximately 6% and 9% HbA1c. Each drug was added in two defined concentrations with concentration one being several times (typically five times) the maximum daily dosage and concentration 2 being the maximum daily dosage level. Concentration one was performed for screening purposes only and concentration two was the relevant drug concentration for determining interferences with the assay. Samples were measured in ten-fold using the cobas c 503 analytical unit. The median value was compared to the reference value (HbA1c sample with no drug added) and the deviation from the reference was calculated. Drug interference studies were conducted using the Hemolysate Application and were representative of both Hemolysate and Whole Blood Applications.
- . N-Acetylcysteine
- Acetylsalicylic acid .
- Ampicillin-Na .
- Ascorbic acid .
- Cefoxitin .
- Heparin .
- Levodopa .
- Methyldopa + 1.5 .
- Metronidazole .
- Doxycyclin .
- Rifampicin .
- Gammagard .
- Cyclosporine .
- Phenylbutazone .
- Acetaminophen .
- Ibuprofen .
- Theophylline .
- Tolbutamide .
Sample Matrix Comparison 4.8.
The purpose of this study was to evaluate Hemoglobin A1c determination, with the Tina-quant Hemoglobin A1cDx Gen.3 assay, in the presence of anticoagulants.
At least 40 samples of each anticoagulant and at least 40 half-filled tubes of each anticoagulant were evaluated. The filled and corresponding half-filled (double concentrated) sample tubes were from one donor. Matrix comparison studies were conducted using the Hemolysate Application and was representative of both Hemolysate and Whole Blood Applications.
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| Sample Type | Anticoagulant | Tube Fill | MeanDifference | Upper 95% | Lower 95% |
|---|---|---|---|---|---|
| Hemolysate | K2-EDTA | ½ Full | 0.004 | 0.124 | -0.116 |
| K3-EDTA | Full | -0.005 | 0.104 | -0.114 | |
| ½ Full | 0.004 | 0.140 | -0.131 | ||
| Na Heparin | Full | 0.000 | 0.120 | -0.121 | |
| ½ Full | -0.026 | 0.129 | -0.181 | ||
| Li Heparin | Full | -0.003 | 0.081 | -0.088 | |
| ½ Full | -0.015 | 0.138 | -0.167 | ||
| NaF/Potassiumoxalate | Full | 0.009 | 0.123 | -0.105 | |
| ½ Full | 0.019 | 0.135 | -0.097 | ||
| EDTA/Fluoride | Full | 0.003 | 0.100 | -0.094 | |
| ½ Full | 0.017 | 0.172 | -0.139 |
Table 18: Matrix Comparison Results
4.9. Method Comparison
A method comparison study was performed to compare the sample results from the candidate method, Hemoglobin A1cDx Gen.3 assay on the cobas c 503 analytical unit, to results from Tosoh Automated Glycohemoglobin Analyzer HLC-723G8 (Tosoh), the NGSP method. This study was conducted with both the Tina-quant Hemoglobin A1cDx Gen.3 Whole Blood Application and Hemolysate Application.
One hundred and seventy-one whole blood samples and one hundred seventy-three hemolysate samples from the secondary NGSP reference laboratory were used in the evaluation. These samples were measured by the secondary NGSP reference laboratory using the Tosoh HPLC system (X-axis) and the Roche Tina-quant Hemoglobin A1cDx Gen.3 test system (Y-axis). The samples were tested over a 3-day period, with one lot of reagent, on one cobas c 503 analytical unit.
| # Samples Tested | % Samples Tested | |||
|---|---|---|---|---|
| % HbA1c | Whole Blood | Hemolysate | Whole Blood | Hemolysate |
| ≤ 5% | 6 | 6 | 3.5% | 3.5% |
| 5-6 % | 23 | 24 | 13.5% | 13.9% |
Table 19: Method Comparison Sample Distribution
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| >6 - 6.5% | 31 | 32 | 18.1% | 18.5% |
|---|---|---|---|---|
| >6.5 - 7% | 37 | 37 | 21.6% | 21.4% |
| >7 – 8% | 25 | 25 | 14.6% | 14.5% |
| >8 – 9% | 13 | 13 | 7.6% | 7.5% |
| >9% | 36 | 36 | 21.1% | 20.8% |
| Total | 171 | 173 | 100% | 100% |
The method comparison demonstrated good agreement between Roche Tina-quant Hemoglobin A1cDx Gen.3 assay and the NGSP Tosoh reference method. The tables below summarize the bias between the Tina-quant Hemoglobin A1cDx Gen.3 assay and the NGSP Tosoh reference method.
Table 20: Difference Plot Analysis Data Summary
| Whole Blood Application | Hemolysate Application | |
|---|---|---|
| Mean bias vs. NGSP TOSOH | -0.046% | 0.046% |
| Mean bias at lower 95% CI | -0.410% | -0.338% |
| Mean bias at upper 95% CI | 0.318% | 0.431% |
Table 21: Bias at Concentration Data Summary
| % HbA1c | % Relative BiasWhole Blood Application | % Relative BiasHemolysate Application |
|---|---|---|
| 5% | -2.4% | 0.6% |
| 6.5% | -1.2% | 0.8% |
| 8% | -0.4% | 1.0% |
| 12% | 0.7% | 1.2% |
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4.10. Total Error
Using the results of bias estimation (%Bias) generated during the method comparison study and precision estimates (%CV) from the precision study, the Total Error (TE) was calculated using the following equation:
%TE = |%Bias| + 1.96 * %CV * (1 + %Bias/100).
Table 22: Total Error – Hemolysate Application
| %HbA1c | % BIAS | Precision (%CV) | Total Error (%) |
|---|---|---|---|
| 4.96 | 0.63 | 1.2 | 3.0 |
| 6.59 | 0.87 | 1.2 | 3.3 |
| 8.29 | 1.02 | 1.3 | 3.5 |
| 12.1 | 1.20 | 1.6 | 4.4 |
Table 23: Total Error – Whole Blood Application
| %HbA1c | % BIAS | Precision (%CV) | Total Error (%) |
|---|---|---|---|
| 4.87 | -2.56 | 1.3 | 5.2 |
| 6.57 | -1.12 | 1.4 | 3.8 |
| 8.20 | -0.30 | 1.4 | 3.1 |
| 12.3 | 0.79 | 1.6 | 3.9 |
ADDITIONAL INFORMATION 5.
Other Devices Marketed with This Assay 5.1.
The following devices are required, but not provided:
- C.f.a.s. (Calibrator for automated systems) HbA1c, K052101 .
- PreciControl HbA1c norm, K103099 .
- PreciControl HbA1c path, K103099 .
CONCLUSIONS 6.
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.