Diazyme 1,5-anhydroglucitol (1,5-AG) Assay is an enzymatic method intended for the quantitative determination of 1,5anhydroglucitol (1,5-AG) in serum or plasma. The 1,5-AG Assay is for the intermediate term (preceding 1-2 weeks) monitoring of glycemic control in people with diabetes. For in vitro diagnostic use only.

Device Description

Diazyme's 1.5-AG assay is an enzymatic method intended for the quantitative determination of 1,5-anhydroglucitol (1,5-AG) in serum or plasma. The assay uses the enzyme pyranose oxidase (PROD) to oxidize the 2nd position hydroxyl group of 1,5-AG and to detect the generated hydrogen peroxide by colorimetry using peroxidase (POD). To eliminate reactive glucose in sample, it is pretreated by enzymatic reactions using hexokinase and pyruvate kinase (PK). Hexokinase uses adenosine triphosphate (ATP) to convert glucose into non-reactive glucose-6-phosphate (G-6-P), generating adenosine diphosphate (ADP). The reaction is driven to completion with PK, as ADP is phosphoralated to ATP during the conversion of phosphoenolpyruvate (PEP) into pyruvate.

AI/ML Overview

The provided document is a 510(k) premarket notification for the Diazyme 1,5-AG Assay, a device for in vitro quantitative determination of 1,5-anhydroglucitol (1,5-AG) in serum or plasma to monitor glycemic control in people with diabetes.

Here's an analysis of the acceptance criteria and the study that proves the device meets them, based on the provided text:

Key Takeaways:

This submission focuses on substantial equivalence to a predicate device (GlycoMark™ 1,5-anhydroglucitol (1,5-AG) K031604) rather than a novel, de novo AI/ML device. Therefore, many standard AI/ML specific criteria like MRMC studies, human-in-the-loop performance, and expert consensus for ground truth are not applicable or detailed in this context.
The studies presented are primarily analytical performance studies to demonstrate that the new device performs comparably to the predicate device and meets established laboratory testing standards.

1. Table of Acceptance Criteria and Reported Device Performance:

The document implicitly defines acceptance by comparing the Diazyme 1,5-AG Assay's performance to the predicate device and established CLSI (Clinical and Laboratory Standards Institute) guidelines for analytical validation.

Acceptance Criteria (Implied)	Predicate Device Performance (GlycoMark™ 1,5-AG)	Reported Device Performance (Diazyme 1,5-AG Assay)
Indications for Use Equivalence	Same as Diazyme 1,5-AG Assay	Same as GlycoMark™ 1,5-AG Assay: Quantitative determination of 1,5-anhydroglucitol (1,5-AG) in serum or plasma for intermediate term (preceding 1-2 weeks) monitoring of glycemic control in people with diabetes. For in vitro diagnostic use only.
Assay Principle Equivalence	Pyranose oxidase (PROD) enzyme, colorimetry	Same: Uses pyranose oxidase (PROD) to oxidize 1,5-AG and detect generated hydrogen peroxide by colorimetry using peroxidase (POD). Pretreatment with hexokinase and pyruvate kinase (PK) to eliminate reactive glucose.
Test Objective Equivalence	Quantitative determination of 1,5-AG	Same: For the in vitro quantitative determination of 1,5-AG in human serum or plasma.
Type of Test Equivalence	Quantitative	Same: Quantitative
Specimen Type Equivalence	Human serum or plasma	Same: Human serum or plasma.
Product Type Equivalence	Assay reagent kit: liquid stable two reagent system	Same: Assay reagent kit: liquid stable two reagent system
Analytical Linearity/Reportable Range	Linear up to 110 µg/mL	Linear up to 116.7 µg/mL. Also linear in the clinically relevant range of 0.6-10.0 µg/mL. (Based on CLSI EP6-A). The recovered values showed good agreement with expected values, with errors generally between 0% and -8%.
Analytical Precision (CV%)	0.8 – 3.8% (2 controls and 2 serum samples)	CV% less than 5% across various samples and runs. (Based on CLSI EP5-A2). Within-run CV% ranged from 0.5% to 2.3%. Total CV% ranged from 0.9% to 4.8%.
Accuracy (Method Comparison)	89.6% concordance (vs. Roche Tina-Quant A1c)	Strong correlation ($R^2$ = 0.9995) with the predicate GlycoMark™ 1,5-AG assay. Slope = 1.0164, Y-intercept = -0.2042. (Based on CLSI EP9-A2). Correlation Coefficient (R) = 0.9997. Standard Error of Estimate = 0.51. The rationale states "excellent correlation."
Limit of Blank (LoB)	Not specified	0.3 µg/mL (Based on CLSI EP17-A2).
Limit of Detection (LoD)	Not specified	0.5 µg/mL (Based on CLSI EP17-A2).
Limit of Quantitation (LoQ)	Not specified	0.6 µg/mL (Based on CLSI EP17-A2).
Analytical Specificity (Interference)	Not specified	Less than 10% deviation caused by: Free Bilirubin (5 mg/dL), Bilirubin Conjugated (5 mg/dL), Hemoglobin (125 mg/dL), Ascorbic Acid (37.5 mg/dL), Triglyceride (1000 mg/dL), Glucose (1000 mg/dL), Maltose (500 mg/dL), Uric Acid (20 mg/dL), Creatinine (10 mg/dL), Urea (20 mg/dL). (Based on CLSI EP7-A2).
Reference Range (Normal Population)	Not specified	Males: 8.19 to 32.19 µg/mL (Non-parametric 5th-95th percentiles). Females: 6.00 to 29.10 µg/mL (Non-parametric 5th-95th percentiles). (Based on CLSI C28-A3 guideline). Note: This is a study result not a performance claim compared to predicate.

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

Precision Study: Six serum samples (tested repeatedly over 240 measurements for each sample to calculate different CV% components).
Linearity Study: Samples encompassing a wide range of 1,5-AG concentrations (Level 0 to Level 10), tested in triplicate for each level. The exact number of distinct patient samples is not specified, but it implies a spiked matrix or dilutions of real samples.
Method Comparison (Accuracy) Study: 102 patient samples covering the Analytical Measurement Range (AMR).
Reference Range Study: 140 apparently healthy males and 140 apparently healthy females (Total 280 samples).
Data Provenance: The document does not specify the country of origin of the data or whether the studies were retrospective or prospective. It refers to "clinical patient samples" for the method comparison study and "apparently healthy males and females" for the reference range study, implying real human samples. Given the nature of in vitro diagnostic device validation, these are typically prospective collection campaigns or use of well-characterized biobank samples.

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

This device is an in vitro diagnostic assay for quantitative measurement of a biomarker, not an AI/ML device interpreting medical images or other complex data. Therefore, the concept of "experts establishing ground truth" in the same way as, for example, radiologists marking tumors, is not applicable.
The "ground truth" for the test set is established by the measurement results obtained from the predicate device (GlycoMark™ 1,5-AG assay) for the method comparison study, and by the inherent concentration values of the samples used in precision, linearity, LoB/LoD/LoQ, and interference studies. These are not dependent on expert visual review or interpretation.
The predicate device itself was validated and approved by the FDA based on its own clinical and analytical performance.

4. Adjudication Method for the Test Set:

Not applicable in this context. Adjudication methods like 2+1 or 3+1 are used when human experts disagree on interpretations (e.g., in radiology image reading). For quantitative in vitro diagnostic assays, the "truth" is the measured value, and deviations are assessed statistically (e.g., bias, correlation, CV%).

5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance:

No, an MRMC comparative effectiveness study was not done. This is an in vitro diagnostic device measuring a chemical biomarker, not an AI system assisting human readers of medical images or other data. The performance is assessed analytically against a predicate device and established laboratory standards.

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

Yes, in essence, the Diazyme 1,5-AG Assay is a standalone device. It performs the quantitative measurement of 1,5-AG in a sample without direct human interpretation of its internal "algorithm" or sensing process for each test. The results are provided as a numeric value. Humans (lab technicians, clinicians) then interpret these numeric results in the clinical context, similar to any lab test. The performance studies (precision, linearity, method comparison, etc.) directly evaluate the performance of this "standalone" assay.

7. The Type of Ground Truth Used (expert consensus, pathology, outcomes data, etc.):

Analytical Reference Values / Predicate Device Measurements:
- For the method comparison study (accuracy), the "ground truth" was the measurements obtained from the legally marketed predicate device (GlycoMark™ 1,5-AG).
- For linearity, LoB/LoD/LoQ, and precision studies, the "ground truth" refers to the expected or true concentrations of the analytes in the reference materials or spiked samples. These values are typically established through highly accurate reference methods or certified reference materials traceable to international standards (though not explicitly detailed, this is standard practice for IVD validation).

8. The Sample Size for the Training Set:

Not applicable in the typical AI/ML sense. This is a chemical assay, not a machine learning algorithm that learns from training data. The "training" for such a device involves optimizing reagents and reaction conditions during development, but there isn't a "training set" of patient data in the same way an AI model is trained.

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

Not applicable. As explained above, this device does not have a "training set" in the context of AI/ML. The "ground truth" for the analytical studies are the established reference values or predicate device measurements.

Summary

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Image /page/0/Picture/0 description: The image contains the logos of the Department of Health and Human Services and the Food and Drug Administration (FDA). The Department of Health and Human Services logo is on the left, and the FDA logo is on the right. The FDA logo includes the text "FDA U.S. FOOD & DRUG ADMINISTRATION" in blue.

October 4, 2018

Diazyme Laboratories Inc. Abhijit Datta VP of Operations 12889 Gregg Court Poway, CA 92130

Re: K180209

Trade/Device Name: Diazyme 1.5-AG Assay Regulation Number: 21 CFR 864.7470 Regulation Name: Glycosylated hemoglobin assay Regulatory Class: Class II Product Code: NOZ Dated: August 23, 2018 Received: August 24, 2018

Dear Abhijit Datta:

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food. Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part

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801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/CombinationProducts/GuidanceRegulatoryInformation/ucm597488.html; good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4. Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm.

For comprehensive regulatory information about medical devices and radiation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/MedicalDevices/DeviceRegulationandGuidance/) and CDRH Learn (http://www.fda.gov/Training/CDRHLearn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website

(http://www.fda.gov/DICE) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).

Sincerely.

Kellie B. Kelm -S

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

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Indications for Use

510(k) Number (if known) K180209

Device Name Diazyme 1,5-AG Assay

Indications for Use (Describe)

Type of Use (Select one or both, as applicable)
☑ Prescription Use (Part 21 CFR 801 Subpart D)	☐ Over-The-Counter Use (21 CFR 801 Subpart C)

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510(k) SUMMARY

This summary of 510(k) safety and effectiveness information is being submitted in accordance
with the requirements of SMDA 1990 and 21 CFR 807.92 The assigned 510(k) number K180209.

Submitter's name:	Diazyme Laboratories Inc.
Submitter's address:	12889 Gregg CourtPoway, CA 92064
Name of Contact Person:	Dr. Abhijit DattaDiazyme Laboratories Inc.12889 Gregg CourtPoway, CA 92064Phone: 858-455-4762
Date the Summary was Prepared:	September 28, 2013
Name of the Device	Diazyme 1,5-AG Assay
Trade Name:	Diazyme 1,5-AG Assay
Common/Usual Name	1,5-anhydroglucitol(1,5-AG) Assay
Device Classification	Assay, Glycosylated Hemoglobin; 21 CFR 864.7470
Product code:	NOZ
Panel:	Hematology (81)
Submission Type	510k
Regulation Number	21 CFR 864.7470
Device Class	II
Predicate Device:	GlycoMarkTM 1,5-anhydroglucitol (1,5-AG) (K031604).
Manufacturing Address	Diazyme Laboratories Inc.12889 Gregg CourtPoway, CA 92064USA
Establishment Registration	2032900

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DESCRIPTION OF THE DEVICE

INDICATIONS FOR USE

Diazyme 1,5-anhydroglucitol (1,5-AG) Assay is an enzymatic method intended for the quantitative determination of 1,5-anhydroglucitol (1,5-AG) in serum or plasma. The 1,5-AG Assay is for the intermediate term (preceding 1-2 weeks) monitoring of glycemic control in people with diabetes. For in vitro diagnostic use only.

Substantial Equivalence Information

Predicate device name(s): GlycoMark™ 1,5-anhydroglucitol (1,5-AG) K031604

Comparison of new device to predicate: The tables below identify similarities and differences between the predicate device and the Diazyme 1,5-AG Assay (K180209).

Indications for Use

GlycoMark™ 1,5-anhydroglucitol (1,5-AG) (K031604)	Diazyme 1,5-AG AssayK180209	Equivalency
The GlycoMark™ test provides quantita-tive measurement of 1,5-anhydroglucitol(15AG) in serum or plasma. The test is forprofessional use, and is indicated for theintermediate term monitoring of glycemiccontrol in people with diabetes.	Diazyme 1,5-AG Assay is an enzymaticmethod intended for the quantitative de-termination of 1,5-anhydroglucitol (1,5-AG) in serum or plasma. The 1,5-AG As-say is for the intermediate term (preced-ing 1-2 weeks) monitoring of glycemiccontrol in people with diabetes. For invitro diagnostic use only.	Same

Assay Principle

GlycoMark™ 1,5-anhydroglucitol(1,5-AG) (K031604)	Diazyme 1,5-AG AssayK180209	Equivalency
The method uses the enzyme pyranose oxidase (PROD) to oxidize the 2nd posi-	Diazyme's 1,5-AG assay uses the enzyme pyranose oxidase (PROD) to oxidize the 2nd position hydroxyl group of 1,5-AG	same

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tion hydroxyl group of 15AG and to de- and to detect the generated hydrogen peroxide by
tect the generated hydrogen peroxide by oxide by colorimetry using peroxidase
colorimetry using peroxidase (POD). As (POD). To eliminate reactive glucose in
PROD reacts with glucose, the sample is sample, it is pretreated by enzymatic reac-
pretreated by enzyme reaction using glu- tions using hexokinase and pyruvate ki-
cokinase (GK). Glucose is converted nase (PK). Hexokinase uses adenosine tri-
into glucose-6-phosphate (G-6-P), a spe- phosphate (ATP) to convert glucose into
cies non-reactive with PROD. To drive non-reactive glucose-6-phosphate (G-6-
the reaction to completion, an adenosine P), generating adenosine diphosphate
triphosphate (ATP)-regenerating system (ADP). The reaction is driven to comple-
consisting of pyruvate kinase (PK) and tion with PK, as ADP is phosphoralated to
phosphoenol pyruvate (PEP) is utilized. ATP during the conversion of phosphoe-
As ATP is converted to adenosine di- nolpyruvate (PEP) into pyruvate.
phosphate (ADP), PK, in the presence of
PEP, catalyzes the phosphorylation of
ADP back to ATP. Following the con-
version of glucose to G-6-P, the assay is
rendered specific for 15AG.

Test Objective

GlycoMark™ 1,5-anhydroglucitol(1,5-AG) (K031604)	Diazyme 1,5-AG AssayK180209	Equivalency
For the in vitro quantitative determina-tion of 1,5-AG in human serum orplasma.	For the in vitro quantitative determina-tion of 1,5-AG in human serum orplasma.	Same

Type of Test

GlycoMark™ 1,5-anhydroglucitol(1,5-AG) (K031604)	Diazyme 1,5-AG AssayK180209	Equivalency
Quantitative	Quantitative	Same

Specimen Type

GlycoMark™ 1,5-anhydroglucitol(1,5-AG) (K031604)	Diazyme 1,5-AG AssayK180209	Equivalency
Human serum or plasma.	Human serum or plasma.	Same

Product Type

Product Type
GlycoMark™ 1,5-anhydroglucitol(1,5-AG) (K031604)	Diazyme 1,5-AG AssayK180209	Equivalency

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Assay reagent kit: liquid stable two rea-gent system	Assay reagent kit: liquid stable two rea-gent system	Same
----------------------------------------------------------	----------------------------------------------------------	------

Performance

GlycoMarkTM 1,5-anhydroglucitol(1,5-AG) (K031604)	Diazyme 1,5-AG AssayK180209
Linearity: up to 110 μg/mL	Linearity: up to 110 μg/mL
Precision: CV% of 0.8 – 3.8% (2 controls and 2 serum samples)	Precision: CV% of less than 5%
Method comparison (vs Roche Tina-Quant A1c):89.6% concordance	Accuracy (vs. GlycoMarkTM 1,5-AG):Correlation Coefficient ( $R^2$ ) = 0.9995, slope = 1.0164, and y intercept = -0.2042.

Rationale for Considering the Device Substantially Equivalent to Devices Approved for Inter State Commerce

GlycoMark™ 1,5-anhydroglucitol (1,5-AG) (K031604) was selected for method comparison with Diazyme 1,5-AG Assay. The method comparison study between the subject and predicate using clinical patient samples showed excellent correlation. The similarities and differences between the predicate reagent and the Diazyme 1,5-AG Assay are given in the table above. Detailed performance characteristics and comparison analysis are given in the filing and demonstrate substantial equivalence to predicate device. The performance characteristics of the Diazyme 1,5-AG Assay are substantially similar to that of the approved predicate test. Performance data and risk analysis indicates that differences should not affect the safety and effectiveness of the Diazyme 1,5-AG Assay and offers users an in vitro diagnostic device system to measure 1.5-AG in human serum and plasma.

Conclusion

Detailed method comparison analysis (accuracy studies) presented in this 510k submission, together with linearity, precision and interference studies, demonstrates that the Diazyme 1,5-AG Assay performance is acceptable, safe and effective. There is no significant deviation between the results obtained by Diazyme 1,5-AG Assay and the legally marketed predicate device GlycoMark™ 1.5-anhydroglucitol (1.5-AG) (K031604) when testing clinical patient samples and is thus substantially similar.

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PERFORMANCE SUMMARY

Precision Study

In the study, six serum samples were tested following CLSI EP5-A2 protocol.

Sample	Meanµg/mL(N=240)	Within-RunSDCV%	Between-RunSDCV%	Between-DaySDCV%	Between-LotSDCV%	TotalSDCV%
S1	3.07	0.072.3%	0.072.3%	0.113.5%	0.144.7%	0.154.8%
S2	5.70	0.071.3%	0.081.4%	0.071.2%	0.132.2%	0.132.2%
S3	11.56	0.090.8%	0.111.0%	0.050.5%	0.151.3%	0.151.3%
S4	23.17	0.140.6%	0.150.6%	0.070.3%	0.220.9%	0.220.9%
S5	61.84	0.320.5%	0.360.6%	0.420.7%	0.631.0%	0.641.0%
S6	97.26	0.480.5%	0.510.5%	0.620.6%	0.931.0%	0.941.0%

Result:

Linearity/Reportable Range

To establish the linearity of the Diazyme 1,5-AG assay, a study design was used based on the CLSI protocol EP6-A: Evaluation of the Linearity of Quantitative Measurement Procedures: a Statistical Approach: Approved Guideline.

Dilution	Recovery Triplicate	Recovery Triplicate	Recovery Triplicate	Recovery Mean (µg/mL)	Expected Recovery (µg/mL)	Error	%Recovery
Level 0	0.2	0.2	0.2	0.20	0.20	0%	100%
Level 1	10.9	10.9	11.0	10.93	11.85	-8%	92%
Level 2	21.8	22.2	22.4	22.13	23.49	-6%	94%
Level 3	34.4	34.2	34.5	34.37	35.14	-2%	98%
Level 4	45.3	45.6	45.0	45.30	46.79	-3%	97%
Level 5	58.4	58.7	58.5	58.53	58.43	0%	100%
Level 6	68.6	68.2	68.9	68.57	70.08	-2%	98%
Level 7	80.7	81.5	81.2	81.13	81.73	-1%	99%
Level 8	92.5	92.7	92.9	92.70	93.37	-1%	99%
Level 9	103.9	103.5	104.5	103.97	105.02	-1%	99%
Level 10	116.7	116.7	116.6	116.67	116.67	0%	100%

Result:

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Dilution	Recovery Triplicate			RecoveryMean (µg/mL)	ExpectedRecovery(µg/mL)	Error	%Recovery
Level 0	0.1	0.1	0.3	0.17	0.17	0%	100%
Level 1	1.4	1.3	1.4	1.37	1.37	0%	100%
Level 2	2.3	2.4	2.5	2.40	2.57	-6%	94%
Level 3	3.5	3.6	3.4	3.50	3.77	-7%	93%
Level 4	4.7	4.6	4.8	4.70	4.97	-5%	95%
Level 5	5.7	5.8	5.8	5.77	6.17	-6%	94%
Level 6	7.0	7.1	7.0	7.03	7.37	-5%	95%
Level 7	8.3	8.2	8.2	8.23	8.57	-4%	96%
Level 8	9.4	9.5	9.6	9.50	9.77	-3%	97%
Level 9	10.9	10.9	10.8	10.87	10.97	-1%	99%
Level 10	12.2	12.2	12.1	12.17	12.17	0%	100%

The Diazyme 1,5-AG Assay is linear up to 116.7 µg/mL, and also in the clinically relevant range of 0.6- 10.0 µg/mL 1.5-AG.

LoB/LoD/LoQ

The Limit of Blank (LoB), the Limit of Detection (LoD) and the Limit of Quantitation (LoQ) of the Diazyme 1,5-AG assay were determined according to CLSI EP17-A2: Protocols for Determination of Limits of Detection and Limits of Quantitation; Approved Guideline using three lots of the reagents on Beckman AU680. The LOB was determined to be 0.3 µg/mL; the LOD was determined to be 0.5 µg/mL; the LOQ was determined to be 0.6 µg/mL.

Analytical specificity

To determine the level of interference from the substances normally present in the patient samples, Diazyme 1,5-AG Assay was used to test low (5.0 µg/mL), medium (23.0 µg/mL), and high (50 ug/mL) 1,5-AG samples with various concentrations of substances following CLSI EP7-A2.

The following substances normally present in the samples produced less than 10% deviation when tested at levels equal to the concentrations listed below.

Interference Substances	Concentration
Free Bilirubin	5 mg/dL
Bilirubin Conjugated	5 mg/dL
Hemoglobin	125 mg/dL
Ascorbic Acid	37.5 mg/dL
Triglyceride	1000 mg/dL
Glucose	1000 mg/dL
Maltose	500 mg/dL
Uric Acid	20 mg/dL
Creatinine	10 mg/dL
Urea	20 mg/dL

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Comparison Studies

Protocol: Clinical and Laboratory Standards Institute EP9-A2 - Method Comparison and Bias Estimation Using Patient Samples: Approved Guideline-Second Edition (2002). For method comparison, the 1,5- AG Assay was evaluated by testing individual serum samples with comparison to predicate legally marketed 1,5- AG device. A total of 102 patient samples covering the AMR were tested by both 1,5- AG Assay and Predicate 1,5- AG Assay.

Result:

Parameter	Results
Slope	1.0164
95% CI	1.012 to 1.021
Intercept	-0.21
95% CI	-0.35 to -0.07
Standard Error of Estimate	0.51
Correlation Coefficient(R)	0.9997
R2	0.9995

Reference Range Study

To determine the reference interval of the normal population, serum samples from 140 apparently healthy males and 140 apparently healthy females were tested using the 1,5- AG Assay according to CLSI C28-A3 guideline.

Using non-parametric 5th 95th percentiles, the reference interval for males was established to be 8.19 to 32.19 µg/mL and 6.00 to 29.10 µg/mL for females.

Regulation Number and Section

§ 864.7470 Glycosylated hemoglobin assay.

(a)
Identification. A glycosylated hemoglobin assay is a device used to measure the glycosylated hemoglobins (A1a , A1b , and A1c ) in a patient's blood by a column chromatographic procedure. Measurement of glycosylated hemoglobin is used to assess the level of control of a patient's diabetes and to determine the proper insulin dosage for a patient. Elevated levels of glycosylated hemoglobin indicate uncontrolled diabetes in a patient.(b)
Classification. Class II (performance standards).