VARIANT II TURBO HbA1c Kit - 2.0

K142448 · Bio-Rad Laboratories, Inc. · PDJ · Mar 24, 2015 · Clinical Chemistry

Device Facts

• Record ID: K142448
• Device Name: VARIANT II TURBO HbA1c Kit - 2.0
• Applicant: Bio-Rad Laboratories, Inc.
• Product Code: PDJ · Clinical Chemistry
• Decision Date: Mar 24, 2015
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 862.1373
• Device Class: Class 2

Intended 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 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 Story

The VARIANT™ II TURBO HbA1c Kit- 2.0 is an in vitro diagnostic assay for professional use in clinical laboratories. It utilizes ion-exchange high-performance liquid chromatography (HPLC) to separate hemoglobins in human whole blood samples. Samples are automatically diluted by the VARIANT™ II TURBO Sampling Station (VSS) and injected into an analytical cartridge. A programmed buffer gradient of increasing ionic strength separates hemoglobins based on ionic interactions. Separated hemoglobins pass through a filter photometer measuring absorbance at 415 nm, with 690 nm used for background correction. The Clinical Data Management (CDM™) software processes raw data, calculates HbA1c area using an exponentially modified Gaussian (EMG) algorithm, and generates reports. Results are used by clinicians to monitor long-term glycemic control and diagnose diabetes. The system benefits patients by providing standardized, precise HbA1c measurements traceable to DCCT and IFCC reference methods.

Clinical Evidence

Bench testing only. Precision evaluated per CLSI EP05-A2 (n=720 measurements per sample). Linearity demonstrated across 3.4–20.6% HbA1c range (R²=0.998). Method comparison (n=130) against NGSP Secondary Reference Laboratory showed Deming slope 1.033 and intercept -0.275. Interference studies confirmed no significant impact from endogenous substances, drugs, or hemoglobin variants (S, C, E, D, A2, F). Total Error calculated at 2.6%–5.2% across clinical decision points.

Technological Characteristics

Ion-exchange HPLC; dual-pump buffer gradient system; filter photometer (415 nm/690 nm); analytical cartridge; automated sampling station. Software-based data reduction and peak identification. Standardized to NGSP/DCCT/IFCC. Professional use only.

Indications for Use

Indicated for quantitative HbA1c determination in human whole blood to aid in diabetes diagnosis and risk assessment. Not for use in newborns, pregnant women, or patients with Type 1 diabetes. Contraindicated for patients with conditions altering red blood cell lifespan (e.g., hemolytic anemia, blood loss, transfusion, hemoglobinopathies, thalassemias), malignancies, or severe hepatic/renal disease. Not for diagnosing gestational diabetes.

Regulatory Classification

Identification

A hemoglobin A1c test system is a device used to measure the percentage concentration of hemoglobin A1c in blood. Measurement of hemoglobin A1c is used as an aid in the diagnosis of diabetes mellitus and as an aid in the identification of patients at risk for developing diabetes mellitus.

Special Controls

(b) Classification. Class II (special controls). Hemoglobin A1c test systems must comply with the following special controls: 1) The device must have initial and annual standardization verification by a certifying glycohemoglobin standardization organization deemed acceptable by FDA. 2) The premarket notification submission must include performance testing to evaluate precision, accuracy, linearity and interference, including the following: i) Performance testing of device precision must, at a minimum, use blood samples with concentrations near 5.0%, 6.5%, 8.0% and 12% hemoglobin A1c. This testing must evaluate precision over a minimum of 20 days using at least 3 lots of the device and 3 instruments, as applicable. ii) Performance testing of device accuracy must include a minimum of 120 blood samples that span the measuring interval of the new device and compare results of the new device to results of the standardized test method. Results must demonstrate little or no bias versus the standardized method. iii) Total error of the new device must be evaluated using single measurements by the new device compared to results of the standardized test method, and this evaluation must demonstrate a total error less than or equal to 6%. iv) Performance testing must demonstrate that there is little to no interference from common hemoglobin variants, including Hemoglobin C, Hemoglobin D, Hemoglobin E, Hemoglobin A2 and Hemoglobin S. 3) When assay interference from Hemoglobin F or interference with other hemoglobin variants with low frequency in the population is observed, a warning statement must be placed in a black box and must appear in all labeling material for these devices describing the interference and any affected populations.

*Classification.* Class II (special controls). The special controls for this device are:(1) The device must have initial and annual standardization verification by a certifying glycohemoglobin standardization organization deemed acceptable by FDA. (2) The premarket notification submission must include performance testing to evaluate precision, accuracy, linearity, and interference, including the following: (i) Performance testing of device precision must, at a minimum, use blood samples with concentrations near 5.0 percent, 6.5 percent, 8.0 percent, and 12 percent hemoglobin A1c. This testing must evaluate precision over a minimum of 20 days using at least three lots of the device and three instruments, as applicable. (ii) Performance testing of device accuracy must include a minimum of 120 blood samples that span the measuring interval of the device and compare results of the new device to results of a standardized test method. Results must demonstrate little or no bias versus the standardized method. (iii) Total error of the new device must be evaluated using single measurements by the new device compared to results of the standardized test method, and this evaluation must demonstrate a total error less than or equal to 6 percent. (iv) Performance testing must demonstrate that there is little to no interference from common hemoglobin variants, including Hemoglobin C, Hemoglobin D, Hemoglobin E, Hemoglobin A2, and Hemoglobin S. (3) When assay interference from Hemoglobin F or interference with other hemoglobin variants with low frequency in the population is observed, a warning statement must be placed in a black box and must appear in all labeling material for these devices describing the interference and any affected populations.

Predicate Devices

• Roche COBAS INTEGRA 800 Tina-quant HbA1c DX Gen. 2 assay (k121291)

Submission Summary (Full Text)

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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 [phone] or [phone] 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 [phone] or [phone] 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)

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.

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)

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#### FOR FDA USE ONLY

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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: [phone] FAX: [phone] E-Mail: Jackie [e-mail]

#### 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:

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.

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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.

#### 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:<br>VARIANT™ II TURBO<br>HbA1c Kit - 2.0 | Predicate Device:<br>COBAS INTEGRA 800<br>HbA1c DX Gen. 2 | Status | | | | | | |
| Intended Use | Intended for the<br>quantitative determination<br>of hemoglobin A1c (IFCC<br>mmol/mol and NGSP %) | Intended for the<br>quantitative determination<br>of hemoglobin A1c (IFCC<br>mmol/mol and NGSP %) | Same | | | | | | |
| Indications for<br>Use -<br>Diagnosis | This test is to be used as<br>an aid in diagnosis of<br>diabetes and as an aid in<br>identifying patients who<br>may be at risk for<br>developing diabetes. | This test is to be used as<br>an aid in diagnosis of<br>diabetes and as an aid in<br>identifying patients who<br>may be at risk for<br>developing diabetes. | Same | | | | | | |
| Indications for<br>Use -<br>Monitoring | Measurement of<br>hemoglobin A1c is<br>effective in monitoring<br>long-term glycemic<br>control in individuals with<br>diabetes mellitus | Measurement of<br>hemoglobin A1c is<br>effective in monitoring<br>long-term glycemic<br>control in individuals with<br>diabetes mellitus | Same | | | | | | |
| Specimen Type | Human Whole blood | Human Whole blood | Same | | | | | | |
| Matrices | K2-EDTA, K3-EDTA | K2-EDTA, K3-EDTA | Same | | | | | | |
| Measuring<br>Interval | 3.4 to 20.6 % (NSGP)<br>14 – 203 mmol/mol<br>HbA1c (IFCC) | 4.3 - 24.8% (NGSP)<br>23 to 258 mmol/mol<br>HbA1c (IFCC) | Same | | | | | | |
| Method<br>Comparison -<br>Whole Blood | N=130<br>y-intercept = 0.2691<br>Slope = 1.0327<br>R2=0.998 | N=141<br>y-intercept= 0.364<br>Slope = 0.955<br>R2=0.989 | Same | | | | | | |
| Total Precision<br>Results<br>NGSP% | Sample<br>%CV<br>5.1%<br>1.6<br>1.3<br>6.6%<br>1.3<br>7.9%<br>12.1%<br>1.1 | Sample<br>%CV<br>5.25%<br>1.5<br>1.4<br>6.63%<br>8.11%<br>1.4<br>12.09%<br>1.5 | Same | | | | | | |
| Standardization | Traceable to the Diabetes<br>Control and<br>Complications Trial<br>(DCCT) reference<br>method and IFCC.<br>Certified via the National<br>Glycohemoglobin<br>Standardization Program<br>(NGSP) | Traceable to the Diabetes<br>Control and<br>Complications Trial<br>(DCCT) reference<br>method and IFCC.<br>Certified via the National<br>Glycohemoglobin<br>Standardization Program<br>(NGSP) | Same | | | | | | |

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| Table 2: Differences with Predicate | | | |
|-------------------------------------|-----------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------|------------------------------------------------------------------------------------|
| Feature | Candidate Device:<br>VARIANT™ II TURBO<br>HbA1c Kit - 2.0 | Predicate Device:<br>COBAS INTEGRA<br>800 HbA1c DX Gen. 2 | Status |
| Hardware | VARIANT™ II TURBO<br>Hemoglobin Testing<br>System and VARIANT™<br>II TURBO Link<br>Hemoglobin Testing<br>System | Roche COBAS<br>INTEGRA 800<br>analyzer | Technology<br>differences<br>do not raise<br>new safety<br>or efficacy<br>concerns |
| Assay Principle | Ion exchange HPLC | Turbidimetric inhibition<br>immunoassay | Technology<br>differences<br>do not raise<br>new safety<br>or efficacy<br>concerns |
| Additional<br>Matrices | Capillary blood in<br>Hemoglobin Capillary<br>Collection System<br>(HCCS) | KF/Na2- EDTA<br>Na-heparin<br>NF/K-oxalate<br>NF/NA2- EDTA<br>Li-Heparin | Technology<br>differences<br>do not raise<br>new safety<br>or efficacy<br>concerns |

#### 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 |
| Concentration<br>HbA1c (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 | Control<br>1 | Control<br>2 | Patient<br>1 | Patient<br>2 | Patient<br>3 | Patient<br>4 | QC 1 | QC 2 | QC 3 |
|--------------------------------|--------------|--------------|--------------|--------------|--------------|--------------|------|------|------|
| Instrument ID: VART17 | | | | | | | | | |
| Concentration<br>HbA1c (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 | Control<br>1 | Control<br>2 | Patient<br>1 | Patient<br>2 | Patient<br>3 | Patient<br>4 | QC 1 | QC 2 | QC 3 |
| Concentration<br>HbA1c (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 | | | |
| Concentration<br>HbA1c (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.

| Sample<br>Pool | Predicted 1st<br>Order | Predicted 2nd<br>Order | 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)

| Sample<br>Pool | Predicted 1 st<br>Order | Predicted 2nd<br>Order | 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-<br>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 – Decision<br>Level | 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)<br>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 Drug<br>Interferent | Highest Level Tested showing no Significant<br>Interference | |
|-------------------------------|-------------------------------------------------------------|--------------|
| | Conventional (US)<br>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 | | | |
|------------------------------------------------------------|----|--------------------------------|----------------------------------|
| Hemoglobin<br>Variant | n | Range in % Abnormal<br>Variant | Range in %HbA1c<br>Concentration |
| 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

| Hemoglobin<br>Variant | Relative % Bias from Reference Method observed at Low and<br>High Concentrations of HbA1c | Relative % Bias (StDev) for<br>HbA1c ~8.0% |
|-----------------------|-------------------------------------------------------------------------------------------|--------------------------------------------|
| | Relative % Bias (StDev)<br>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 |

- 4. 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 verification<br>by a certifying glycohemoglobin standardization organization<br>deemed acceptable by the FDA. |
| YES | Performance testing of device precision must, at a minimum,<br>use blood samples with concentrations near 5.0%, 6.5%, 8.0% |
| YES | and 12% hemoglobin A1c. Testing must evaluate precision<br>over a minimum of 20 days using at least 3 lots of the device<br>and 3 instruments, as applicable. |
| YES | Performance testing of accuracy must include a minimum of<br>120 blood samples that span the measuring interval of the new<br>device and compare results of the new device to results of the<br>standardized method. Results must demonstrate little or no<br>bias versus the standardized method. |
| YES | Total error of the new device must be evaluated using single<br>measurements by the new device compared to results of the<br>standardized test method, and this evaluation must<br>demonstrate a total error less than or equal to 6%. |
| YES | Performance testing must demonstrate that there is little to no<br>interference from common hemoglobin variants, including<br>Hemoglobin C, Hemoglobin D, Hemoglobin E, Hemoglobin A2<br>and Hemoglobin S. |
| N/A | When assay interference from Hemoglobin F or interference<br>with other hemoglobin variants with low frequency in the<br>population is observed, a warning statement must be placed in<br>a black box and must appear in all labeling material for these<br>devices describing the interference and any affected<br>population. |

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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.