The VARIANT™ II TURBO HbA1c Kit- 2.0 is intended for the quantitative determination of hemoglobin A1c (IFCC mmol/mol and NGSP %) in human whole blood using ion-exchange high-performance liquid chromatography (HPLC) on the VARIANT II TURBO Hemoglobin Testing System and VARIANT II TURBO Link Hemoglobin Testing System.

This test is to be used as an aid in diagnosis of diabetes and as an aid in identifying patients who may be at risk for developing diabetes.

The VARIANT II TURBO HbA1c Kit- 2.0 is intended for Professional Use Only.

The Hemoglobin Capillary Collection System (HCCS) is intended for the collection of human whole blood for the percent determination of hemoglobin A1c using Bio-Rad HPLC methods.

Device Description

The VARIANT™ II TURBO HbA1c Kit - 2.0 utilizes principles of ion-exchange highperformance liquid chromatography (HPLC). The samples are automatically diluted on the VARIANT™ II TURBO Sampling Station (VSS) and injected into the analytical cartridge. The VARIANT™ II TURBO Chromatographic Station (VCS) dual pumps deliver a programmed buffer gradient of increasing ionic strength to the cartridge, where the hemoglobins are separated based on their ionic interactions with the cartridge material. The separated hemoglobins then pass through the flow cell of the filter photometer, where changes in the absorbance at 415 nm are measured. An additional filter at 690 nm corrects for background absorbance.

The VARIANT™ II TURBO Clinical Data Management (CDM™) software performs reduction of raw data collected from each analysis. Two-level calibration is used for adjustment of the calculated HbA1c values. A sample report, including retention times of detected peaks and a chromatogram, is generated by the CDM for each sample. The A1c peak is shaded for ease of identification. The area is calculated using an exponentially modified Gaussian (EMG) algorithm and the result printed in either mmol/mol or % HbA1c format as selected by the user.

The VARIANT™ II TURBO HbA1c Kit – 2.0 is designed to be used on the standalone VARIANT™ II TURBO and the VARIANT™ II TURBO Link Hemoglobin Testing Systems. VARIANT™ II TURBO and the VARIANT™ II TURBO Link Hemoglobin Testing Systems are identical with respect to all operational and system components. Physically, the VARIANT™ II TURBO Link VSS outer case is modified for compatibility with a track Functionality of the VARIANT™ II TURBO Link has not changed, just the system. physical orientation to accommodate sample tube management on a track system.

AI/ML Overview

This document describes the performance of the VARIANT™ II TURBO HbA1c Kit - 2.0, an in vitro diagnostic device used for quantitative determination of hemoglobin A1c.

Here's an analysis of the acceptance criteria and the study that proves the device meets them:

1. Table of Acceptance Criteria and Reported Device Performance

The acceptance criteria are derived from the "Special Controls Requirements for HbA1c for Diabetes Diagnosis" as outlined in Table 17 of the document (though it's mislabeled and actually on page 14 of the PDF). The device performance is reported in various sections of the "Summary of Performance Data."

Requirement (Acceptance Criteria)	Reported Device Performance and Section in Document	Does Device Meet Criteria?
Device must have initial and annual standardization verification by a certifying glycohemoglobin standardization organization deemed acceptable by the FDA.	Section 8.d Traceability: "The assigned HbA1c values... are certified with the National Glycohemoglobin Standardization Program (NGSP)." (p. 10)	Yes
Performance testing of device precision must, at a minimum, use blood samples with concentrations near 5.0%, 6.5%, 8.0%, and 12% hemoglobin A1c. Testing must evaluate precision over a minimum of 20 days using at least 3 lots of the device and 3 instruments, as applicable.	Section 8.a Precision/Reproducibility: Evaluated based on CLSI EP05-A2 using four EDTA whole blood samples at ~5%, ~6.5%, ~8%, and ~12% HbA1c. Precision evaluated using three reagent lots, three instruments, for 20 days. (p. 6)	Yes
Performance testing of accuracy must include a minimum of 120 blood samples that span the measuring interval of the new device and compare results of the new device to results of the standardized method. Results must demonstrate little or no bias versus the standardized method.	Section 8.c Method Comparison: 130 variant-free whole blood EDTA samples (3.4% to 20.0% HbA1c) were evaluated. Deming and Passing-Bablok regression analyses performed against an NGSP SRL reference method. Bias estimation (Table 11) shows low bias. (p. 8-10)	Yes
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%.	Section 8.c Total Error Decision Levels: Total Error (TE) calculated at 5.0%, 6.5%, 8.0%, and 12.0% HbA1c. All reported TE values (5.2%, 3.4%, 2.6%, 3.2% respectively) are less than 6%. (Table 12, p. 10)	Yes
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.	Section 8.e.iv Hemoglobin Variant Study: Specific variant samples (HbS, HbC, HbD, HbE, HbA2, HbF) were tested. Relative % Bias from Reference Method observed for each variant reported in Table 16 (p. 13). All biases are within acceptable limits, indicating little to no interference.	Yes
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 population.	Section 8.e.iv Hemoglobin Variant Study: HbF was included in the study, and its bias was reported. No warning statement recommendation is mentioned, implying no significant interference was observed that would require such a warning. (p. 13)	N/A (No significant interference observed)

Detailed Study Information:

2. Sample Size Used for the Test Set and the Data Provenance

Precision/Reproducibility Study:
- Sample Size: 4 EDTA whole blood samples (at ~5%, ~6.5%, ~8%, and ~12% HbA1c) and 5 quality control materials. Each sample had 720 measurements (run in duplicate, 2 runs/day, for 20 days across 3 instruments).
- Data Provenance: Not explicitly stated regarding country of origin, but it involves patient samples and quality control materials, which are typically retrospective or derived from clinical settings.
Method Comparison Study:
- Sample Size: 130 variant-free whole blood EDTA samples.
- Data Provenance: Not explicitly stated regarding country of origin. The samples ranged from 3.4% to 20.0% HbA1c, suggesting prospective collection covering a diagnostic range. It mentions comparison to a "secondary NGSP SRL reference laboratory," implying real-world clinical samples.
Linearity Study:
- Sample Size: Low (3.4%HbA1c) and high (20.6%HbA1c) EDTA whole blood patient samples mixed at varying ratios, creating 11 distinct levels (Level 1 to Level 11).
- Data Provenance: Patient samples, likely retrospective.
Analytical Specificity (Interference) Studies:
- Endogenous Interference: Two EDTA whole blood sample pools (~6.5%HbA1c and ~8.0%HbA1c) were used. 10 replicates of each pool with test and control samples.
- Drug Interference: Two EDTA whole blood sample pools (~6.5%HbA1c and ~8.0%HbA1c) were used. 10 replicates of each drug with test and control samples.
- Hemoglobin Derivatives Interference: Low (~6.5%HbA1c) and high (~8.0%HbA1c) whole blood EDTA samples. Ten replicates each.
- Hemoglobin Variant Study: Specific variant samples (HbS, HbC, HbD, HbE, HbA2, HbF), with n=21-29 for each variant (e.g., 26 for HbS, 25 for HbC). Tested in duplicate.
- Data Provenance: Not explicitly stated regarding country of origin. "Patient samples" were used for some, and pure substances/serum samples for others.

3. Number of Experts Used to Establish the Ground Truth for the Test Set and the Qualifications of Those Experts

The document does not explicitly state the number or qualifications of experts for establishing ground truth.
For the Method Comparison Study, the device's results were compared to a "secondary NGSP SRL reference laboratory using a cleared HPLC-based HbA1c assay." The NGSP (National Glycohemoglobin Standardization Program) provides certification for laboratories and methods to ensure traceability to the DCCT (Diabetes Control and Complications Trial) reference method. This implies the ground truth is established by a highly standardized and validated reference method in an accredited laboratory, rather than by individual experts' consensus.
For the Hemoglobin Variant Study, the comparison was against "a NGSP reference method that has been demonstrated to be free from the hemoglobin interferent," again pointing to a high-standard reference method as the ground truth.

4. Adjudication Method for the Test Set

No explicit adjudication method (like 2+1 or 3+1 consensus) is described. The ground truth relies on established reference methods (NGSP-certified HPLC for HbA1c, and specialized NGSP reference methods for variant studies) as opposed to human adjudication of ambiguous cases. This is typical for quantitative analytical devices where a measurable reference standard exists.

5. If a Multi Reader Multi Case (MRMC) Comparative Effectiveness Study was Done

No, an MRMC comparative effectiveness study was not done. This type of study typically evaluates the effectiveness of a diagnostic aid (like AI) on physician performance in interpreting medical images or making diagnoses. The VARIANT™ II TURBO HbA1c Kit - 2.0 is an automated in vitro diagnostic device for quantitative chemical analysis, not an image interpretation or diagnostic aid that involves human "readers." Its performance is measured directly against reference methods and statistical criteria.

6. If a Standalone (i.e. algorithm only without human-in-the loop performance) was Done

Yes, the studies evaluate the standalone (algorithm only) performance. The device is an automated system (ion-exchange HPLC) that processes samples and produces quantitative HbA1c results. The performance data presented (precision, linearity, method comparison, interference studies) directly assesses the accuracy and reliability of this automated system in determining HbA1c values, without human intervention in the interpretative step. The "Human-in-the-loop" aspect would primarily involve sample collection and loading the instrument, not interpretation of the results themselves.

7. The Type of Ground Truth Used

Primarily reference method comparison, traceable to international standards.
- For HbA1c quantification, the ground truth is established by comparison to NGSP-certified reference methods, which are themselves traceable to the DCCT (Diabetes Control and Complications Trial) reference method and IFCC (International Federation of Clinical Chemistry and Laboratory Medicine) units. This is stated under "Traceability" (Section 8.d) and in the method comparison and variant studies.
- For interference studies, the ground truth is based on the expected values of spiked samples without the interferent, as measured by the device itself or a control.

8. The Sample Size for the Training Set

The document does not explicitly mention a "training set" in the context of machine learning or algorithm development. This device is based on established HPLC principles and data reduction algorithms, which are typically designed and validated, rather than "trained" in the machine learning sense. The performance studies described serve as validation/verification of the final device.

9. How the Ground Truth for the Training Set Was Established

As a traditional in vitro diagnostic device based on HPLC, there isn't a "training set" in the machine learning paradigm. The device's underlying principles (ion-exchange chromatography, spectrophotometry, and specific algorithms for peak detection and calculation) are built upon scientific understanding and engineering. The "ground truth" for developing and calibrating such systems would come from well-characterized reference materials and methods, likely through extensive internal R&D and calibration procedures following international standards, but this is not detailed as a "training set" in the document.

Summary

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Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002

April 3, 2015

BIO-RAD LABORATORIES, INC. JACKIE BUCKLEY REGULATORY AFFAIRS REP IV 4000 ALFRED NOBEL DR. HERCULES CA 94547

Re: K142448

Trade/Device Name: VARIANT™ II TURBO HbA1c Kit - 2.0 Hemoglobin Capillary Collection System Regulation Number: 21 CFR 862.1373 Regulation Name: Glycosylated hemoglobin assay Regulatory Class: II Product Code: PDJ, JKA Dated: March 24, 2015 Received: March 24, 2015

Dear Ms. Jackie Buckley:

This letter corrects our substantially equivalent letter of March 24, 2015.

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

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CFR Part 807); labeling (21 CFR Parts 801 and 809] ); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

If you desire specific advice for your device on our labeling regulation (21 CFR Part 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" (21CFR 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,

Stayce Beck -S

Courtney H. Lias, Ph.D. For : Director Division of Chemistry and Toxicology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health

Enclosure

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Indications for Use

510(k) Number (if known) K142448

Device Name VARIANT™ II TURBO HbA1c Kit- 2.0

Hemoglobin Capillary Collection System

Indications for Use (Describe)

This test is to be used as an aid in diagnosis of diabetes and as an aid in identifying patients who may be at risk for developing diabetes.

The VARIANT II TURBO HbA1c Kit- 2.0 is intended for Professional Use Only.

The Hemoglobin Capillary Collection System (HCCS) is intended for the collection of human whole blood for the percent determination of hemoglobin A1c using Bio-Rad HPLC methods.

Type of Use (Select one or both, as applicable)

2 Prescription Use (Part 21 CFR 801 Subpart D)

_ Over-The-Counter Use (21 CFR 801 Subpart C)

PLEASE DO NOT WRITE BELOW THIS LINE - CONTINUE ON A SEPARATE PAGE IF NEEDED.

FOR FDA USE ONLY

Concurrence of Center for Devices and Radiological Health (CDRH) (Signature)

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.

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Department of Health and Human Services Food and Druq 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."

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510(k) Summary (Summary of Safety and Effectiveness)

This Summary of 510(k) Safety and Effectiveness is being submitted in accordance with the requirements of SMDA 1990 and 21 CFR 807.92.

The assigned 510(k) number is:

Date Summary prepared: March 20, 2015

1. Applicant Name:

Bio-Rad Laboratories, Inc. Clinical Diagnostics Group 4000 Alfred Nobel Drive Hercules, California 94547

2. Contact Person:

Jackie Buckley Telephone Number: (510) 741-5309 FAX: (510) 741-6471 E-Mail: Jackie buckley@bio-rad.com

3. Device Name/Trade Name:

VARIANT™ II TURBO HbA1c Kit – 2.0 Classification Name: Hemoglobin A1c Test System Common Name: HbA1c Product Code: PDJ C.F.R Section: 21 CFR 862.1373 Device classification: Class II

Hemoglobin Capillary Collection System Classification Name: Tubes, Vials, Systems, Serum Separators, Blood Collection Product Code: JKA C.F.R Section: 21 CFR 862.1675 Device classification: Class II

4. Predicate Device:

COBAS INTEGRA 800 Tina-quant HBA1cDx Gen.2 assay (K121291)

5. Description of the Device:

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6. Indications for Use:

The VARIANT™ II TURBO HbA1c Kit – 2.0 is intended for the quantitative determination of hemoglobin A1c (IFCC mmol/mol and NGSP %) in human whole blood using ionexchange high-performance liquid chromatography (HPLC) on the VARIANT™ II TURBO Hemoglobin Testing System and VARIANT™ II TURBO Link Hemoglobin Testing System.

This test is to be used as an aid in diagnosis of diabetes and as an aid in identifying patients who may be at risk for developing diabetes.

The VARIANT™ II TURBO HbA1c Kit – 2.0 is intended for Professional Use Only.

The Hemoglobin Capillary Collection System (HCCS) is intended for the collection of human whole blood for the percent determination of hemoglobin A1c using Bio-Rad HPLC methods.

7. Substantial Equivalence Information:

Predicate Device Name:

Roche COBAS INTEGRA 800 Tina-quant HbA1c DX Gen. 2 assay

Predicate 510(k) number: K121291

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Table 1: Similarities with Predicate
Feature	Candidate Device:VARIANT™ II TURBOHbA1c Kit - 2.0	Predicate Device:COBAS INTEGRA 800HbA1c DX Gen. 2	Status
Intended Use	Intended for thequantitative determinationof hemoglobin A1c (IFCCmmol/mol and NGSP %)	Intended for thequantitative determinationof hemoglobin A1c (IFCCmmol/mol and NGSP %)	Same
Indications forUse -Diagnosis	This test is to be used asan aid in diagnosis ofdiabetes and as an aid inidentifying patients whomay be at risk fordeveloping diabetes.	This test is to be used asan aid in diagnosis ofdiabetes and as an aid inidentifying patients whomay be at risk fordeveloping diabetes.	Same
Indications forUse -Monitoring	Measurement ofhemoglobin A1c iseffective in monitoringlong-term glycemiccontrol in individuals withdiabetes mellitus	Measurement ofhemoglobin A1c iseffective in monitoringlong-term glycemiccontrol in individuals withdiabetes mellitus	Same
Specimen Type	Human Whole blood	Human Whole blood	Same
Matrices	K2-EDTA, K3-EDTA	K2-EDTA, K3-EDTA	Same
MeasuringInterval	3.4 to 20.6 % (NSGP)14 – 203 mmol/molHbA1c (IFCC)	4.3 - 24.8% (NGSP)23 to 258 mmol/molHbA1c (IFCC)	Same
MethodComparison -Whole Blood	N=130y-intercept = 0.2691Slope = 1.0327R2=0.998	N=141y-intercept= 0.364Slope = 0.955R2=0.989	Same
Total PrecisionResultsNGSP%	Sample%CV5.1%1.61.36.6%1.37.9%12.1%1.1	Sample%CV5.25%1.51.46.63%8.11%1.412.09%1.5	Same
Standardization	Traceable to the DiabetesControl andComplications Trial(DCCT) referencemethod and IFCC.Certified via the NationalGlycohemoglobinStandardization Program(NGSP)	Traceable to the DiabetesControl andComplications Trial(DCCT) referencemethod and IFCC.Certified via the NationalGlycohemoglobinStandardization Program(NGSP)	Same

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Table 2: Differences with Predicate
Feature	Candidate Device:VARIANT™ II TURBOHbA1c Kit - 2.0	Predicate Device:COBAS INTEGRA800 HbA1c DX Gen. 2	Status
Hardware	VARIANT™ II TURBOHemoglobin TestingSystem and VARIANT™II TURBO LinkHemoglobin TestingSystem	Roche COBASINTEGRA 800analyzer	Technologydifferencesdo not raisenew safetyor efficacyconcerns
Assay Principle	Ion exchange HPLC	Turbidimetric inhibitionimmunoassay	Technologydifferencesdo not raisenew safetyor efficacyconcerns
AdditionalMatrices	Capillary blood inHemoglobin CapillaryCollection System(HCCS)	KF/Na2- EDTANa-heparinNF/K-oxalateNF/NA2- EDTALi-Heparin	Technologydifferencesdo not raisenew safetyor efficacyconcerns

8. Summary of Performance Data:

a. Precision/Reproducibility:

The precision of the VARIANT™ II TURBO HbA1c Kit – 2.0 was evaluated based on CLSI EP05-A2 guidelines, Evaluation of Precision Performance of Quantitative Measurement Methods using a modified study design. Four EDTA whole blood samples at the following targeted HbA1c concentrations of ~5%, ~6.5%, ~8% and ~12% were utilized in the study. In addition, five quality control materials were also tested. Precision was evaluated using three reagent lots, three VARIANT™ II TURBO Hemoglobin Testing Systems at two different sites. The samples were run in duplicate in 2 runs per day for 20 days. For each sample, there were 720 measurements. Results are shown in Tables 3-6.

Variation Source	Instrument ID: VART15
	Control 1	Control 2	Patient 1	Patient 2	Patient 3	Patient 4	QC 1	QC 2	QC 3
ConcentrationHbA1c (NGSP%)	5.5	9.9	5.1	6.7	8.0	12.0	5.5	9.9	15.0
Repeatability	0.5%	0.3%	0.5%	0.6%	1.0%	0.3%	0.6%	0.4%	0.4%
Between-Run	0.4%	0.0%	0.3%	0.0%	0.2%	0.3%	0.3%	0.3%	0.2%
Between-Day	0.8%	0.6%	0.8%	0.7%	0.6%	0.5%	1.2%	0.8%	0.6%
Between-Lot	0.8%	0.6%	1.0%	0.8%	0.6%	0.6%	1.0%	0.5%	0.2%
Total Precision	1.4%	0.9%	1.4%	1.2%	1.3%	0.9%	1.7%	1.0%	0.8%

Table 3: Instrument 1 (% CV by Sample (NGSP))

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Variation Source	Control1	Control2	Patient1	Patient2	Patient3	Patient4	QC 1	QC 2	QC 3
Instrument ID: VART17
ConcentrationHbA1c (NGSP%)	5.5	9.8	5.1	6.6	7.9	2.0	5.5	9.9	14.9
Repeatability	0.5%	0.3%	0.6%	0.6%	0.5%	0.4%	0.6%	0.5%	0.4%
Between-Run	0.4%	0.2%	0.5%	0.0%	0.3%	0.4%	0.0%	0.2%	0.3%
Between-Day	0.5%	0.3%	0.4%	0.5%	0.7%	0.3%	0.9%	0.7%	0.4%
Between-Lot	0.9%	0.7%	0.9%	0.7%	0.6%	0.4%	1.3%	0.6%	0.3%
Total Precision	1.2%	0.9%	1.3%	1.0%	1.1%	0.8%	1.7%	1.1%	0.8%

Table 4: Instrument 2 (% CV by Sample (NGSP))

Table 5: Instrument 3 (% CV by Sample (NGSP))

	Instrument ID: VartGerm01
Variation Source	Control1	Control2	Patient1	Patient2	Patient3	Patient4	QC 1	QC 2	QC 3
ConcentrationHbA1c (NGSP%)	5.4	9.7	5.1	6.6	8.0	12.1	5.4	9.8	15.0
Repeatability	0.6%	0.5%	0.8%	0.8%	0.5%	0.4%	0.8%	0.4%	0.4%
Between-Run	0.2%	0.0%	0.1%	0.0%	0.0%	0.2%	0.0%	0.2%	0.0%
Between-Day	0.6%	0.3%	0.6%	0.5%	0.5%	0.4%	0.7%	0.4%	0.3%
Between-Lot	2.0%	0.9%	1.6%	1.4%	1.0%	0.7%	2.2%	1.1%	0.7%
Total Precision	2.2%	1.1%	1.9%	1.7%	1.3%	0.9%	2.5%	1.2%	0.9%

Table 6: Instruments Combined (% CV by Sample (NGSP))

Variation Source	Control 1	Control 2	All Instruments				QC 1	QC 2	QC 3
			Patient 1	Patient 2	Patient 3	Patient 4
ConcentrationHbA1c (NGSP %)	5.4	9.8	5.1	6.6	7.9	12.1	5.4	9.9	15.0
Repeatability	0.5%	0.4%	0.7%	0.7%	0.7%	0.4%	0.7%	0.5%	0.4%
Between-Run	0.3%	0.0%	0.4%	0.0%	0.2%	0.3%	0.2%	0.2%	0.2%
Between-Day	0.7%	0.4%	0.6%	0.5%	0.6%	0.4%	1.0%	0.7%	0.5%
Between-Instrument	1.3%	1.1%	0.4%	0.0%	0.4%	0.6%	0.8%	0.4%	0.0%
Between-Lot	1.4%	0.8%	1.2%	1.0%	0.8%	0.6%	1.6%	0.7%	0.5%
Total Precision	2.1%	1.5%	1.6%	1.3%	1.3%	1.1%	2.2%	1.2%	0.8%

b. Linearity

A linearity study was performed per CLSI EP06-A: Evaluation of the Linearity of Quantitative Measuring Procedures; A Statistical Approach. Linearity across the reportable range was performed using low (3.4%HbA1c) and high (20.6%HbA1c) EDTA whole blood patient samples. These samples were mixed together in varying ratios. The measured values were compared to the theoretical values based upon the dilution factor. Polynomial regression analysis (for first, second, and third order polynomials) were performed to determine the statistical significance of non-linearity. The higher order coefficients were found not to be significant and linearity was demonstrated.

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% HbA1c (NGSP) using the VARIANTTM II TURBO HbA1c Kit – 2.0 has been demonstrated linear from 3.4 – 20.6% HbA1c with the maximum measured difference of ± 0.03% between the predicted 1st and 20 order results as shown in Table 7 below. mmol/mol HbA1c (IFCC) has been demonstrated as linear from 14 – 203 mmol/mol with the maximum measured difference of ± 0.3% (or +/- 0.38mmol/mol) as shown in Table 8 below.

SamplePool	Predicted 1stOrder	Predicted 2ndOrder	Difference
Level 1	3.39	3.41	-0.03
Level 2	5.12	5.13	-0.01
Level 3	6.85	6.85	0.00
Level 4	8.58	8.57	0.01
Level 5	10.31	10.29	0.01
Level 6	12.04	12.02	0.01
Level 7	13.77	13.75	0.01
Level 8	15.50	15.49	0.01
Level 9	17.23	17.23	0.00
Level 10	18.96	18.98	-0.02
Level 11	20.69	20.72	-0.03

Table 7: Results of Linearity Study (NGSP %)

Table 8: Results of Linearity Study (IFCC mmol/mol)

SamplePool	Predicted 1 stOrder	Predicted 2ndOrder	Difference
Level 1	14	14	-0.32
Level 2	32	33	-0.14
Level 3	51	51	-0.01
Level 4	70	70	0.09
Level 5	89	89	0.15
Level 6	108	108	0.16
Level 7	127	127	0.13
Level 8	146	146	0.07
Level 9	165	165	-0.04
Level 10	184	184	-0.19
Level 11	203	203	-0.38

c. Method Comparison
A Method comparison study was performed per CLSI EP09-A2 IR, Method Comparison and Bias Estimation Using Patient Samples. 130 variant-free whole blood EDTA samples ranging from 3.4% to 20.0% HbA1c were evaluated using the VARIANT™ II TURBO HbA1c Kit- 2.0 on the VARIANT™ II TURBO Hemoglobin Testing System. Samples were tested in a single determination over a 4 day period. The results were amples were tested in a single delemination over a 4 day period. The results were
mpared to testing performed at a secogdary NGSP SRL reference laboratory using a
rummary 510(k) S

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cleared HPLC-based HbA1c assay. The distribution of samples spanned the measuring interval listed in Table 9.

Hemoglobin A1c level	n	% Samples tested
≤ 5%	6	4.6
5 - 6%	17	13.1
6 - 6.5%	33	25.4
6.5 - 7%	31	23.8
7 - 8%	21	16.2
8 - 9%	11	8.5
> 9%	11	8.5
Total samples	130	100

Table 9: Distribution of samples

Deming (weighted) and Passing-Bablok regression analyses were performed for the VARIANT™ II TURBO HbA1c Kit - 2.0 versus the NGSP SRL reference method.

Table 10: Summary of Method Comparison Results

	y-Intercept	95% CI	Slope	95% CI
Deming	-0.275	-0.342 – 0.208	1.033	1.023 – 1.043
Passing-Bablok	-0.331	-0.419 – 0.255	1.041	1.029 – 1.054

Image /page/9/Figure/6 description: This image is a scatter plot with a Deming fit. The x-axis is labeled "%HbA1c, NGSP" and ranges from 2 to 22. The y-axis is labeled "% HbA1c, VARIANT II TURBO HbA1c Kit- 2" and ranges from 2 to 22. The data points are clustered along a line, and there is a Deming fit line plotted through the data.

Figure 1: Scatter Plot using Deming Fit, %HbA1c, NGSP SRL vs. VARIANT™ II TURBO HbA1c Kit - 2.0

510(k) Summary

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The following biases between VARIANT™ II TURBO HbA1c Kit – 2.0 versus NGSP SRL Method (Reference method) were observed:

% HbA1c – DecisionLevel	Bias	% Bias
5.0	-0.11	-2.11
6.5	-0.06	-0.87
8.0	-0.01	-0.09
12.0	0.12	1.03

Table 11: Bias Estimation

Total Error Decision Levels

Using the results of bias estimation (%Bias) in the method comparison study and precision estimates in the reproducibility study, Total Error (TE) at four concentrations: (5.0 %, 6.5%, 8.0% and 12.0%) were calculated as follows: %TE=l%Bias| + 1.96 CV (1 + %Bias). The results are presented in the Table 12.

% A1c – Decision Level	% Bias	% CV	% TE
5.0	-2.11	1.6	5.2
6.5	-0.87	1.3	3.4
8.0	-0.09	1.3	2.6
12.0	1.03	1.1	3.2

Table 12: Total Error Estimation

d. Traceability, Stability, Expected Values (calibrators)

The assigned HbA1c values of the VARIANT™ II TURBO HbA1c Kit -2.0 are certified with the National Glycohemoglobin Standardization Program (NGSP). The final reportable result is traceable to both the International Federation of Clinical Chemistry (IFCC) and the Diabetes Control and Complications Trial (DCCT). The International Federation of Clinical Chemistry (IFCC) units of mmol/mol are calculated using the Master Equation NGSP (%) = 0.09148 x IFCC (mmol/mol) + 2.152. HbA1c results are provided to the customers using two different units: NGSP equivalent units (%) and IFCC equivalent units (mmol/mol).

Calibrator Materials:

Value assignment for calibrators (VARIANT™ II TURBO HbA1c Kit - 2.0 Calibrator/Diluent Set) which are recommended for use with this device, were previously reviewed under 510(k) submission K070452.

e. Analytical specificity:

i.) Endogenous Interference
An Endogenous Interference study was performed per CLSI EP07-A2, Interference Testing in Clinical Chemistry. Two EDTA whole blood sample pools were evaluated using a low level whole blood sample with a concentration ~6.5%HbA1c and a high level whole blood sample with a concentration of HbA1c of ~8.0%.

Conjugated bilirubin, unconjugated bilirubin and glucose, available in pure form, were obtained and stock solutions prepared at 10x the intended test

510(k) Summary

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concentration. The 10x stock solution of the test substance was pipetted into a low whole blood sample pool (at ~6.5% HbA1c) and a high whole blood sample pool (~8.0% HbA1c), making the test pool. Ten replicates of each pool prepared with the test and control samples were analyzed using the VARIANT™ II TURBO HbA1c Kit-2.0 on the VARIANT™ II TURBO Hemoglobin Testing System.

Rheumatoid factor, lipemia and total protein were not available as pure standards therefore serum samples with known concentration of these compounds were used. The test pool was prepared by mixing the serum sample known to have a high test substance concentration with a whole blood non-variant sample such that the concentration of test substance in the final mixture would be at the desired level. Ten replicates of each pool prepared with the test and control samples were analyzed using the VARIANT™ II TURBO HbA1c Kit-2.0 on the VARIANT™ II TURBO Hemoqlobin Testing System.

Significant interference was defined as a ± 7% change in %HbA1c value from the control. Results in Table 13 showed no significant interference up to the stated concentrations.

	Concentration
Endogenous substance	Conventional (US)units	SI Units
Lipemia (Intralipid)	6000 mg/dL	60 g/L
Conjugated bilirubin	60 mg/dL	712 µmol/L
Unconjugated bilirubin	60 mg/dL	1026 µmol/L
Glucose	2000 mg/dL	111 mmol/L
Rheumatoid factor	750 IU/mL	750 kIU/mL
Total protein	21 g/dL	210 g/L

Table 13: Endogenous Interference Study Results

ii.) Druq Interference:

A Drug Interference study was performed based per CLSI EP07-A2, Interference Testing in Clinical Chemistry. Two EDTA whole blood sample pools were evaluated using a low level whole blood sample with a concentration ~6.5%HbA1c and a high level whole blood sample with a concentration of ~8.0%HbA1c. Test samples were prepared by spiking each drug at the interferent concentration shown in Table 14. Ten replicates of each drug prepared with the test and control samples were analyzed using the VARIANT™ II TURBO HbA1c Kit-2.0 on the VARIANT™ II TURBO Hemoglobin Testing System.

Significant interference was defined as a more than ± 7% change in %HbA1c value from the control. No significant interference was observed at therapeutic levels up to the stated concentrations in Table 14.

Table 14: Drug Interference Study Results

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Potential DrugInterferent	Highest Level Tested showing no SignificantInterference
	Conventional (US)units	SI units
Acetylcysteine	166 mg/dL	10.2 mmol/L
Ampicillin-Na	1000 mg/dL	28.65 mmol/L
Ascorbic acid	300 mg/dL	17.05 mmol/L
Cefoxitin	2500 mg/dL	58.55 mmol/L
Heparin	5000 U/L	5000 U/L
Levodopa	20 mg/dL	1015 µmol/L
Methyldopa	20 mg/dL	948 µmol/L
Metronidazole	200 mg/dL	11.7 mmol/L
Doxycyclin	50 mg/dL	1124 µmol/L
Acetylsalicylic acid	1000 mg/dL	55.51 mmol/L
Rifampicin	64 mg/L	78 µmol/L
Cyclosporine	5 mg/L	4 µmol/L
Acetaminophen	200 mg/L	1323 µmol/L
Ibuprofen	500 mg/L	2427 µmol/L
Theophylline	100 mg/L	556 µmol/L
Phenylbutazone	400 mg/L	1299 µmol/L

iii.) Cross Reactivity with Hemoglobin Derivatives:

A Hemoglobin Derivatives Interference study was performed based on CLSI EP07-A2, Interference Testing in Clinical Chemistry. Potential interference from Acetylated hemogloblin (Hb), Carbamylated hemoglobin (Hb) and Labile HbA1c were evaluated using a low level whole blood EDTA sample with a concentration ~6.5%HbA1c and a high level whole blood EDTA sample with a concentration of ~8.0% HbA1c. The potentially interfering hemoglobin derivatives were spiked into the low and high level blood samples and each sample was analyzed using ten replicates each in the same analytical run on the VARIANT™ II TURBO Hemoglobin Testing System with the VARIANT™ II TURBO HbA1c Kit - 2.0.

510(k) Summary

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Significant interference was defined as more than a ±7% change in HbA1c value from the control. The test result conclusions are as follows:

Acetylated Hb- up to 50 mg/dL does not interfere with this assay. •
Carbamylated Hb up to 4% (2.6 mM potassium cyanate) does not . interfere with this assay.
. Labile A1c- up to 6% (1000mg/dL) glucose does not interfere with this assay.

Results showed there was no cross reactivity with these substances at physiological levels.

iv.) Hemoglobin Variant Study:

A Hemoglobin Variant study was performed using specific variant samples known to contain hemoglobin variants S, C, E, D, A2 and F. Two whole blood EDTA patient samples containing an HbA1c ~6.5% and ~ 8% and the appropriate hemoglobin variant were tested. Testing of the samples containing hemoglobin variants S, C, E, D , A2 and F were performed in duplicate. Testing of the samples was performed using the VARIANT™ II TURBO HbA1c Kit – 2.0 on the VARIANT™ II TURBO Hemoglobin Testing System and compared to results obtained by a NGSP reference method that has been demonstrated to be free from the hemoglobin interferent. Table 15 contains the number of samples, range of samples and concentration of samples used in the Hemoglobin Variant Study. Table 16 contains the results for the Hemoglobin Variant study bias.

Table 15: Variant samples used in Hemoglobin Variant Study
HemoglobinVariant	n	Range in % AbnormalVariant	Range in %HbA1cConcentration
HbS	26	26.8 - 41.6	6.0 - 8.6
HbC	25	33.3 - 42.4	6.1 - 7.9
HbD	21	30.2 - 41.9	6.1 - 8.6
HbE	24	24.7 - 31.4	6.1 - 8.3
HbA2	22	5.0 - 10.2	5.4 - 14.5
HbF	29	3.5 - 29.2	5.4 - 14.4

d in Homoglobin Vorignt Studi Table 15: Variant complac use

Table 16: Hemoqlobin Variant Study Bias Results

HemoglobinVariant	Relative % Bias from Reference Method observed at Low andHigh Concentrations of HbA1c	Relative % Bias (StDev) forHbA1c ~8.0%
	Relative % Bias (StDev)for HbA1c ~6.5%
HbS	1.9 (± 2.8)	2.8 (± 1.8)
HbC	-0.3 (± 3.5)	-2.5 (±- 2.5)
HbD	-1.1 (± 1.7)	-1.2 (±- 1.0)
HbE	0.7 (± 3.0)	2.2 (± 1.4)
HbF	-1.9 (±- 3.1)	-0.1 (±-2.1)
HbA2	1.4 (± 2.3)	2.0 (±4.1)

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2. Matrix comparison

The data supports the use of the following blood collection tubes and collection system with the VARIANT™ II TURBO HbA1c Kit – 2.0.

. K2-EDTA
. K3-EDTA
Hemoglobin Capillary Collection System (HCCS). .

3. Expected Values/Reference Range

Hemoglobin A1c expected values range was cited from American Diabetes Association Standards of Medical Care in Diabetes 2010, 33 (Supplement 1), S62-S69 for Diagnosis of Diabetes.

Table 17: Hemoglobin A1c Expected Values

Hemoglobin A1c		Suggested Diagnosis
NGSP %	IFCC mmol/mol
>6.5	>47	Diabetic
5.7 — 6.4	39-46	Pre-Diabetic
<5.7	<39	Non-Diabetic

1. Satisfaction of Special Controls Requirements for Diabetes Diagnosis Claim

VARIANT™ II TURBO/TURBO Link Status of Special Controls for HbA1c for Diabetes Diagnosis

Yes/No	Requirement
YES	Device must have initial and annual standardization verificationby a certifying glycohemoglobin standardization organizationdeemed acceptable by the FDA.
YES	Performance testing of device precision must, at a minimum,use blood samples with concentrations near 5.0%, 6.5%, 8.0%
YES	and 12% hemoglobin A1c. Testing must evaluate precisionover a minimum of 20 days using at least 3 lots of the deviceand 3 instruments, as applicable.
YES	Performance testing of accuracy must include a minimum of120 blood samples that span the measuring interval of the newdevice and compare results of the new device to results of thestandardized method. Results must demonstrate little or nobias versus the standardized method.
YES	Total error of the new device must be evaluated using singlemeasurements by the new device compared to results of thestandardized test method, and this evaluation mustdemonstrate a total error less than or equal to 6%.
YES	Performance testing must demonstrate that there is little to nointerference from common hemoglobin variants, includingHemoglobin C, Hemoglobin D, Hemoglobin E, Hemoglobin A2and Hemoglobin S.
N/A	When assay interference from Hemoglobin F or interferencewith other hemoglobin variants with low frequency in thepopulation is observed, a warning statement must be placed ina black box and must appear in all labeling material for thesedevices describing the interference and any affectedpopulation.

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Conclusion:

The information and data in this 510(k) document demonstrate that the VARIANT™ II TURBO HbA1c Kit - 2.0 as performed on the VARIANT™ II TURBO and VARIANT™ II TURBO Link Hemoglobin Testing Systems is an accurate, reliable, precise test that correlates well with current cleared methods and NGSP standardized testing for the quantitation of HbA1c. The contents of this submission demonstrates that the VARIANT™ II TURBO HbA1c Kit – 2.0 as performed on the VARIANT™ II TURBO and VARIANT™ II TURBO Link Hemoglobin Testing Systems is substantially equivalent to its predicate device, Cobas Integra 800 Tina-quant HbA1cDX Gen. 2 Assay and, therefore, safe and effective for its intended use. The performance criteria as stipulated by the Special Controls requirements for HbA1c systems that diagnose diabetes have clearly been met. The VARIANT™ II TURBO HbA1c Kit - 2.0 must be found to be substantially equivalent to the predicate and, therefore, cleared by the agency for the intended use requested.

Regulation Number and Section

§ 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.