Predicate Devices

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The description focuses on standard HPLC principles and data processing using algorithms like EMG, which are not indicative of AI/ML. There is no mention of AI, ML, or related terms in the document.

Therapeutic

No
This device is for the quantitative determination of hemoglobin A1c to aid in the diagnosis of diabetes and identify patients at risk, which is a diagnostic purpose, not a therapeutic one.

Diagnostic

Yes

The device is intended for the "quantitative determination of hemoglobin A1c" and "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." This explicitly states its role in diagnosis.

SaMD (Software as a Medical Device)

No

The device is a kit used with a physical HPLC system and includes hardware components (analytical cartridge, buffers, etc.) for performing the test. While it includes software for data processing, it is not solely software.

IVD (In Vitro Diagnostic)

Yes, this device is an IVD (In Vitro Diagnostic).

Here's why:

Intended Use: The intended use explicitly states "quantitative determination of hemoglobin A1c... in human whole blood". This is a measurement performed on a biological sample (whole blood) taken from the human body.
Purpose: The test is used "as an aid in diagnosis of diabetes and as an aid in identifying patients who may be at risk for developing diabetes." This indicates a medical purpose related to diagnosing or assessing a medical condition.
Device Description: The device description details a system that analyzes the components of the blood sample using chemical and physical principles (ion-exchange high-performance liquid chromatography, photometer). This is characteristic of an in vitro diagnostic device.
Professional Use Only: The indication for "Professional Use Only" is common for IVD devices, as they are intended for use by trained healthcare professionals in a laboratory or clinical setting.

Therefore, based on the provided information, the VARIANT™ II TURBO HbA1c Kit- 2.0 and its associated systems are clearly intended for in vitro diagnostic use.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / 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 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.

Product codes (comma separated list FDA assigned to the subject device)

PDJ, JKA

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 (CDMTM) 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.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

Not Found

Indicated Patient Age Range

Not Found

Intended User / Care Setting

Professional Use Only.

Description of the training set, sample size, data source, and annotation protocol

Not Found

Description of the test set, sample size, data source, and annotation protocol

Not Found

Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)

a. Precision/Reproducibility:
Study Type: Precision/Reproducibility.
Sample Size: Seven hundred twenty measurements were taken for each of four whole blood samples (HbA1c concentrations of ~5%, ~6.5%, ~8%, ~12%) and five quality control materials.
Data Source: Four EDTA whole blood samples and five quality control materials.
Key Results: 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. Results were presented as % CV by sample (NGSP) for each instrument and combined instruments. Total Precision ranged from 0.8% to 2.5% across different samples and controls.

b. Linearity:
Study Type: Linearity.
Sample Size: Not explicitly stated, but involved mixing low (3.4% HbA1c) and high (20.6% HbA1c) EDTA whole blood patient samples in varying ratios.
Key Results: Linearity was demonstrated for % HbA1c (NGSP) from 3.4% to 20.6% with a maximum measured difference of ±0.03% between predicted 1st and 2nd order results. Linearity was demonstrated for mmol/mol HbA1c (IFCC) from 14 to 203 mmol/mol with a maximum measured difference of ±0.3% (or +/- 0.38 mmol/mol).

c. Method Comparison:
Study Type: Method Comparison.
Sample Size: 130 variant-free whole blood EDTA samples.
Data Source: Samples ranging from 3.4% to 20.0% HbA1c, evaluated using the VARIANT™ II TURBO HbA1c Kit- 2.0 on the VARIANT™ II TURBO Hemoglobin Testing System. Compared to testing performed at a secondary NGSP SRL reference laboratory using a cleared HPLC-based HbA1c assay.
Key Results: Deming (weighted) and Passing-Bablok regression analyses were performed.
Deming: y-intercept = -0.275 (95% CI: -0.342 – 0.208), Slope = 1.033 (95% CI: 1.023 – 1.043).
Passing-Bablok: y-intercept = -0.331 (95% CI: -0.419 – 0.255), Slope = 1.041 (95% CI: 1.029 – 1.054).
Bias estimation showed biases ranging from -0.11% at 5.0% HbA1c to 0.12% at 12.0% HbA1c. Total Error (TE) ranged from 2.6% to 5.2% across different decision levels.

d. Traceability, Stability, Expected Values (calibrators):
Key Results: Assigned HbA1c values traceable to NGSP, IFCC, and DCCT. Expected values cited from American Diabetes Association Standards of Medical Care in Diabetes 2010.

e. Analytical specificity:
i.) Endogenous Interference:
Study Type: Endogenous Interference.
Key Results: No significant interference (defined as ±7% change) was observed from Lipemia (Intralipid), Conjugated bilirubin, Unconjugated bilirubin, Glucose, Rheumatoid factor, and Total protein at stated concentrations.

ii.) Drug Interference:
Study Type: Drug Interference.
Key Results: No significant interference (defined as more than ±7% change) was observed from various drugs (e.g., Acetylcysteine, Ampicillin-Na, Ascorbic acid) at therapeutic levels up to the stated concentrations.

iii.) Cross Reactivity with Hemoglobin Derivatives:
Study Type: Hemoglobin Derivatives Interference.
Key Results: No significant interference (defined as more than ±7% change) was observed from Acetylated Hb (up to 50 mg/dL), Carbamylated Hb (up to 4% / 2.6 mM potassium cyanate), and Labile HbA1c (up to 6% / 1000 mg/dL glucose) at physiological levels.

iv.) Hemoglobin Variant Study:
Study Type: Hemoglobin Variant Study.
Sample Size: HbS (n=26), HbC (n=25), HbD (n=21), HbE (n=24), HbA2 (n=22), HbF (n=29).
Key Results: For common hemoglobin variants (S, C, D, E, F, A2), the relative % Bias from the reference method was generally low, indicating minimal interference. For example, HbS showed a bias of 1.9% (±2.8) at ~6.5% HbA1c and 2.8% (±1.8) at ~8.0% HbA1c.

Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)

Not Found. Key metrics reported are different:
Precsion: %CV
Accuracy/Bias: %Bias, %TE

Predicate Device(s): If the device was cleared using the 510(k) pathway, identify the Predicate Device(s) K/DEN number used to claim substantial equivalence and list them here in a comma separated list exactly as they appear in the text. List the primary predicate first in the list.

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

Reference Device(s): Identify the Reference Device(s) K/DEN number and list them here in a comma separated list exactly as they appear in the text.

Not Found

Predetermined Change Control Plan (PCCP) - All Relevant Information for the subject device only (e.g. presence / absence, what scope was granted / cleared under the PCCP, any restrictions, etc).

Not Found

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.

Summary

0

Image /page/0/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo features a stylized depiction of an eagle or bird with three profiles of human faces incorporated into its design. The words "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" are arranged in a circular pattern around the bird symbol.

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

1

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

2

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)

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.

The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to:

Department of Health and Human Services Food and 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."

3

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:

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.

4

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

5

Table 1: Similarities with Predicate
Feature	Candidate Device:
VARIANT™ II TURBO
HbA1c Kit - 2.0	Predicate Device:
COBAS INTEGRA 800
HbA1c DX Gen. 2	Status
Intended Use	Intended for the
quantitative determination
of hemoglobin A1c (IFCC
mmol/mol and NGSP %)	Intended for the
quantitative determination
of hemoglobin A1c (IFCC
mmol/mol and NGSP %)	Same
Indications for
Use -
Diagnosis	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.	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.	Same
Indications for
Use -
Monitoring	Measurement of
hemoglobin A1c is
effective in monitoring
long-term glycemic
control in individuals with
diabetes mellitus	Measurement of
hemoglobin A1c is
effective in monitoring
long-term glycemic
control in individuals with
diabetes mellitus	Same
Specimen Type	Human Whole blood	Human Whole blood	Same
Matrices	K2-EDTA, K3-EDTA	K2-EDTA, K3-EDTA	Same
Measuring
Interval	3.4 to 20.6 % (NSGP)
14 – 203 mmol/mol
HbA1c (IFCC)	4.3 - 24.8% (NGSP)
23 to 258 mmol/mol
HbA1c (IFCC)	Same
Method
Comparison -
Whole Blood	N=130
y-intercept = 0.2691
Slope = 1.0327
R2=0.998	N=141
y-intercept= 0.364
Slope = 0.955
R2=0.989	Same
Total Precision
Results
NGSP%	Sample
%CV
5.1%
1.6
1.3
6.6%
1.3
7.9%
12.1%
1.1	Sample
%CV
5.25%
1.5
1.4
6.63%
8.11%
1.4
12.09%
1.5	Same
Standardization	Traceable to the Diabetes
Control and
Complications Trial
(DCCT) reference
method and IFCC.
Certified via the National
Glycohemoglobin
Standardization Program
(NGSP)	Traceable to the Diabetes
Control and
Complications Trial
(DCCT) reference
method and IFCC.
Certified via the National
Glycohemoglobin
Standardization Program
(NGSP)	Same

6

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

7

| Variation Source | Control
1 | Control
2 | Patient
1 | Patient
2 | Patient
3 | Patient
4 | QC 1 | QC 2 | QC 3 |
|--------------------------------|--------------|--------------|--------------|--------------|--------------|--------------|------|------|------|
| Instrument ID: VART17 | | | | | | | | | |
| Concentration
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
1	Control
2	Patient
1	Patient
2	Patient
3	Patient
4	QC 1	QC 2	QC 3
Concentration
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
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.

8

% 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
Pool | Predicted 1st
Order | Predicted 2nd
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
Pool | Predicted 1 st
Order | Predicted 2nd
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

9

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

10

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

| % HbA1c – Decision

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

11

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

12

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

13

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
Variant	n	Range in % Abnormal
Variant	Range in %HbA1c
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
Variant | Relative % Bias from Reference Method observed at Low and
High Concentrations of HbA1c | Relative % Bias (StDev) for
HbA1c ~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) |

14

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