DATA PACK Catalog #: H7546

DP-H7546-01 04/14 Page 2 of 9

About the New Product

The new Pointe Scientific two part HbA1c assay (H7546) is a stabilized modification of the previous, innovative HbA1c three part reagent that was first introduced in 2003. The improved reagent maintains the same antibodies and buffers, but includes additional stabilizers in the R2 to allow it to become a single component as opposed to two separate components that required mixing prior to use.

Background

Throughout the circulatory life of the red cell, Hemoglobin A1c is formed continuously by the adduction of glucose to the N-terminal of the hemoglobin beta chain. This process, which is non-enzymatic, reflects the average exposure of hemoglobin to glucose over an extended period. In a classical study, Trivelli et al1 showed Hemoglobin A1c in diabetic subjects to be elevated 2-3 fold over the levels found in normal individuals. Several investigators have recommended that Hemoglobin A1c serve as an indicator of metabolic control of the diabetic, since Hemoglobin A1c levels approach normal values for diabetics in metabolic control.2,3,4

Hemoglobin A1c has been defined operationally as the “fast fraction” hemoglobins (HbA1a, A1b, A1c) that elute first during column chromatography with cation-exchange resins. The non-glycosylated hemoglobin, which consists of the bulk of the hemoglobin has been designated HbA0. The present procedure utilizes a antigen and antibody reaction to directly determine the concentration of the HbA1c.

Summary and Explanation of Test

This method utilizes the interaction of antigen and antibody to directly determine the HbA1c in whole blood. Total hemoglobin and HbA1c have the same unspecific absorption rate to latex particles. When mouse antihuman HbA1c monoclonal antibody is added (R2), latex-HbA1c-mouse anti human HbA1c antibody complex is formed. Agglutination is formed when goat anti-mouse IgG polyclonal antibody interacts with the monoclonal antibody. The amount of agglutination is proportional to the amount of HbA1c absorbed on to the surface of latex particles. The amount of agglutination is measured as absorbance. The HbA1c value is obtained from a calibration curve.

DATA PACK Catalog #: H7546

DP-H7546-01 04/14 Page 3 of 9

Equivalence to Pointe Scientific, Inc. Predicate Device:

Parameter New Two Part HbA1c (H7546) Pointe Scientific, Inc. (H7541)

Intended use Same

For the quantitative determination of Hemoglobin A1c (HbA1c) in

human blood. The determination of HbA1c is most commonly

performed for the evaluation of glycemic control in diabetes

mellitus. HbA1c values provide an indication of glucose levels over

the preceding 4-8 weeks. A higher HbA1c value indicates poorer glycemic control. For in vitro

diagnostic use only. Specimen type Same Whole Blood

Detection Same Immunoturbidimetric

Reagents

2 Reagents R1 – Latex particles in a buffered solution with stabilizers R2 – Single reagent with a monoclonal Ab against human HbA1c and polyclonal anti- mouse Ab in buffered solution with stabilizers

3 Reagents R1 – Latex particles in a buffered solution with stabilizers R2a and R2b – Mouse anti-human HbA1c monoclonal antibody, goat anti-mouse IgG polyclonal antibody, Buffer, stabilizers.

Storage conditions Same Stored at 2-8oC Assay range Same 2.0%-16.0%

Interference Same

No interference (≤ ± 10%) from bilirubin 50 mg/dL Ascorbic acid 50mg/dL Triglycerides 2000 mg/dL Carbamylated Hb 7.5mmol/L Acetylated Hb 5.0 mmol/L Same

Specificity Same Hemoglobin variants HbA2, HbC and HbS do not interfere with this method

Sensitivity A 0.056 absorbance change is approximately equivalent to 1.0% HbA1c.

A 0.073 absorbance change is approximately equivalent to 1.0% HbA1c.

Precision Within run %CV: 0.8 % to 2.3 % Day to Day %CV: 1.1 % to 3.2 %

Within run %CV 1.4 % to 1.7 % Day to Day %CV: 2.7 % to 2.8 %

DATA PACK Catalog #: H7546

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Performance Summary

Parameter Output Sample Type(s) Whole Blood Predicate Method/Product Pointe Scientific, Inc. HbA1c Three Part (H7541)

Comparison

Correlation 0.995

Slope 0.989

Intercept -0.047

Samples (n) 40

Precision

Levels Low

Med

High

Within Day Low – 2.3 %

Med – 0.8 %

High – 1.2%

Day to Day Low – 3.2 %

Med – 1.1 %

High – 1.5 %

Shelf Life >18 months (accelerated) Linear Range 0.83 – 17.93 %

Sensitivity Change in absorbance of 0.056 absorbance change for 1% HbA1c

DATA PACK Catalog #: H7546

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Regulatory and Certifications: CE mark applied: Yes 510(k): K031539 Clearance Date: 08/06/2003 NGSP Certification Date: October 1, 2013 IFCC Certificate Date: December 5, 2013 CLIA categorization:

TEST SYSTEM COMPLEXITY EFFECTIVE DATE

Shenzhen Mindray Bio-Medical BS-200 Chemistry Analyzer MODERATE 1/5/2009 Roche Diagnostics Hitachi 917 MODERATE 6/23/2005 Abbott Aeroset MODERATE 4/26/2005 Abbott Alcyon MODERATE 4/26/2005 Bayer Technicon RA 1000 MODERATE 4/26/2005 Bayer Technicon RA 500 MODERATE 4/26/2005 Bayer Technicon RA XT MODERATE 4/26/2005 Beckman Coulter Synchron CX 4 MODERATE 4/26/2005 Beckman Coulter Synchron CX 5 MODERATE 4/26/2005 Bio-Chem Laboratory Systems ATAC 8000 MODERATE 4/26/2005 Ciba Corning 550 Express MODERATE 4/26/2005 Olympus AU 400 MODERATE 4/26/2005 Olympus AU 600 MODERATE 4/26/2005 Roche Diagnostics COBAS MIRA MODERATE 4/26/2005 Roche Diagnostics Cobas Mira Plus MODERATE 4/26/2005 Roche Diagnostics Hitachi 704 MODERATE 4/26/2005 Roche Diagnostics Hitachi 717 MODERATE 8/06/2003 Roche Diagnostics Hitachi 902 MODERATE 4/26/2005 Roche Diagnostics Hitachi 911 MODERATE 4/26/2005 Roche Diagnostics Hitachi 917 MODERATE 4/26/2005 Vitalab Selectra E MODERATE 4/26/2005 Stanbio Sirrus Clinical Chemistry Analyzer MODERATE 2/08/2007

Sizes Available: 40ml kit (R1 30 mL/ R2 10 mL)

120ml kit (R1 90 mL/ R2 30 mL)

Bulk format (3:1 ratio)

DATA PACK Catalog #: H7546

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Instrument Applications Available: • Abbot Aeroset • Beckman/Olympus AU400/640 • Beckman CX • Beckman DxC • Beckman LX • Biolis 24i • Pentra 400 • Vitros 5 • Pointe 360 • Randox Daytona • Cobas 6000 c501 • Cobas Mira • Hitachi 717 • Hitachi 911 • Hitachi 917 • Bayer Advia 1650/1800 • Bayer Advia 2400 • Dimension • Selectra SE • ATAC 8000 • Alycon • Biotecnica BT2000/3000 • Chemwell • Cobas Mira Plus • Envoy 500 • Express • Hitachi 902 • Hitachi 704 • Hitachi 747 • Hitachi 912 • Hitachi 914 • Konelab 20 • Konelab 30/60 • Mindray BS200 • Mindray BS300 • Mindray BS400 • Beckman Olympus AU2700/5400 • Prestige 24i • RA500-1000 • Sirrus • Spectrum Instruments • Targa * Bolded items have validated applications

DATA PACK Catalog #: H7546

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Associated Products: Calibrators: 4 level Calibrator Set (H7541-CAL) Controls: 2 Level Control Set (H7541-CTL)/ Normal and Elevated

Method Comparison:

4

5

6

7

8

9

10

11

4 5 6 7 8 9 10 11

Existing

New

Correlation analysis

X axis (H7541)Existing Y axis (H7546) New

n 40 r 0.995

Slope 0.989 Intercept -0.047

DATA PACK Catalog #: H7546

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Precision: Within-Day precision

Statistics Low Med High

n 20 20 20 Mean 5.970 8.490 12.210

SD 0.138 0.072 0.152 CV 2.31% 0.85% 1.24%

Data

Sample Normal Med High 1 5.90 8.50 12.10 2 5.60 8.50 12.00 3 5.90 8.50 12.20 4 5.90 8.50 12.30 5 5.90 8.50 12.30 6 5.90 8.50 12.00 7 5.90 8.50 12.30 8 6.00 8.50 12.60 9 6.20 8.50 12.30

10 5.90 8.50 12.20 11 6.00 8.60 12.20 12 5.90 8.60 12.30 13 5.90 8.40 11.90 14 6.10 8.50 12.20 15 6.00 8.60 12.40 16 6.20 8.40 12.20 17 6.00 8.40 12.20 18 6.00 8.50 12.10 19 6.00 8.30 12.20 20 6.20 8.50 12.20

Day-to-Day Precision Statistics Low Med High

n 20 20 20 Mean 5.945 8.335 12.150

SD 0.190 0.093 0.179 CV 3.20% 1.12% 1.47%

Data

Sample Normal Med High 1 6.10 8.40 12.20 2 6.00 8.40 12.20 3 5.90 8.40 12.10 4 5.90 8.30 12.00 5 5.80 8.20 12.50 6 5.90 8.20 12.30 7 5.80 8.20 12.20 8 5.90 8.30 12.40 9 5.90 8.40 12.00

10 5.80 8.40 12.00 11 6.00 8.30 12.00 12 5.90 8.40 11.70 13 6.40 8.40 12.10 14 6.00 8.50 12.10 15 5.80 8.30 12.10 16 6.10 8.50 12.10 17 6.40 8.30 12.10 18 5.90 8.20 12.40 19 5.70 8.30 12.30 20 5.70 8.30 12.20

DATA PACK Catalog #: H7546

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Linearity:

0

5

10

15

20

0 5 10 15 20

Theoretical value (%)

Mea

sure

d va

lue(

%)

(Shelf Life):

References: 1. Trivelli, L.A., Ranney, H.M., and Lai, H.T., New Eng. J. Med. 284,353 (1971). 2. Gonen, B., and Rubenstein, A.H., Diabetologia 15, 1 (1978). 3. Gabbay, K.H., Hasty, K., Breslow, J.L., Ellison, R.C., Bunn, H.F., and Gallop, P.M., J. Clin.

Endocrinol. Metab. 44, 859 (1977). 4. Bates, H.M., Lab. Mang., Vol 16 (Jan. 1978).

Theoretical Measured Recovery[%] [%] [%]

0.00 0.00 1.70 0.83 48.8%5.60 4.47 79.8%

11.10 10.23 92.2%17.60 17.57 99.8%

Controls target T=0 7 M 12 M 20 M 25 M

Level 1 4.2 – 6.2 5.9 5.6 5.6 5.6 5.7 Recovery - 95% 95% 95% 97%

Level 2 10.0 – 13.0 11.7 11.7 12 12.00 12.3

Recovery - 100% 103% 103% 105% unit: [%Hb]

Acceptance criteria: Original value + 10%