N-GENEOUS HDL CHOLESTEROL KIT/CHOLESTEROL CALIBRATOR by GENZYME CORP.

The Genzyme N-geneous™ HDL Cholesterol Kit, is a two-reagent homogenous method for use in the direct quantitative determination of high density lipoprotein cholesterol (HDL-C) in human serum and plasma. The first readent contains a mixture of polymers and polvanions that bind to the sufface of low-density lipoproteins (LDL), very low-density lipoproteins (VLDL) and chylomicrons. These complexed lipoproteins are stabilized, even in the presence of detergent which is added as part of the second reagent, together with cholesterol enzymes. HDL particles, on the other hand, are not stabilized by the polymers and polvanions and become solubilized by the detergent. Consequently, only the HDL cholesterol is subject to cholesterol measurement.

AI/ML Overview

Here's an analysis of the provided text, focusing on the acceptance criteria and the study used to prove the device meets these criteria:

1. Table of Acceptance Criteria and Reported Device Performance:

The document doesn't explicitly state quantitative acceptance criteria in a dedicated section. However, based on the comparative performance studies, the implicit acceptance criteria for equivalence seem to be a high correlation coefficient and a regression equation demonstrating close agreement with the predicate methods. For precision, the acceptance criteria would be low %CV values, indicating good reproducibility.

Performance Metric	Acceptance Criteria (Implicit)	Reported Device Performance (N-geneous™ HDL Cholesterol Kit)
Comparative Performance
Correlation Coefficient (r) vs. Phosphotungstic Acid	High (e.g., >0.90, typically >0.95 for good agreement)	0.96
Regression Equation vs. Phosphotungstic Acid	Slope close to 1, intercept close to 0	y = 0.81x + 7.82
Correlation Coefficient (r) vs. CDC Method	High (e.g., >0.90, typically >0.95 for good agreement)	0.96
Regression Equation vs. CDC Method	Slope close to 1, intercept close to 0	y = 1.01x - 3.39
Precision (Within-Run)
%CV (Low HDL)	Low %CV (e.g., <5%)	1.50%
%CV (Mid HDL)	Low %CV (e.g., <5%)	1.18%
%CV (High HDL)	Low %CV (e.g., <5%)	1.04%
Precision (Between-Run)
%CV (Low HDL)	Low %CV (e.g., <5%)	3.79%
%CV (Mid HDL)	Low %CV (e.g., <5%)	2.64%
%CV (High HDL)	Low %CV (e.g., <5%)	2.78%

The conclusion states, "These data demonstrate that the performance of the N-geneous™ HDL Cholesterol Kit in the clinical laboratory is substantially equivalent to the performance of the PTA and CDC Reference methods." This implies that the reported performance met the acceptance criteria for substantial equivalence.

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

Comparative Studies (N-geneous™ vs. Phosphotungstic Acid):
- Sample Size: 108 serum samples
- Data Provenance: The document states "patient serum samples with HDL values between 27-91 mg/dL (5th and 95th percentively) were used for these comparative studies." This suggests the samples were from a clinical population. The studies were conducted at "three sites," implying real-world clinical settings. The data is retrospective as it uses pre-existing or collected patient samples. The country of origin is not explicitly stated.
Comparative Studies (N-geneous™ vs. CDC Reference Method):
- Sample Size: A subset of at least 25 serum samples (specifically, "n = 26" is reported in the table).
- Data Provenance: Same as above – patient serum samples from the clinical population, likely retrospective, country of origin not specified.
Precision Studies:
- Sample Size: For within-run and between-run studies, 3 target levels (low, mid, high) were used.
  - Within-run: n=20 for each level (total 60 measurements on different aliquots of the same pool).
  - Between-run: n=80 for each level (total 240 measurements on different aliquots of the same pool over time).
- Data Provenance: "fresh and frozen serum pools" were used. These are typically prepared in a laboratory setting. The country of origin is not specified.

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

Not Applicable (N/A) / No Experts Used in this Context: For an in-vitro diagnostic (IVD) device like an HDL Cholesterol Kit, "ground truth" is established by a reference method or a predicate device, not by human expert interpretation in the same way it would be for imaging or clinical diagnosis.
- The "experts" in this context are the established, validated laboratory methods themselves: the Phosphotungstic Acid MgCl (PTA) precipitation method and the Center for Disease Control (CDC) reference method for HDL-C.

4. Adjudication Method for the Test Set:

Not Applicable (N/A): Adjudication, like 2+1 or 3+1, is typically used when human interpretation contributes to establishing ground truth for ambiguous cases, often in imaging studies. For an IVD, the reference methods provide definite numerical results, so no human adjudication of results is necessary. The comparison is entirely quantitative.

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: This is an in-vitro diagnostic (IVD) device, not an AI-powered diagnostic imaging or clinical decision support tool that would involve human readers or AI assistance. Therefore, an MRMC study and analysis of human reader improvement with AI are not relevant or applicable here.

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

Yes, by its nature: The "Genzyme N-geneous™ HDL Cholesterol Kit" is a standalone assay. It generates a numerical result (HDL-C concentration) directly from a serum/plasma sample without human interpretation or intervention in the measurement process itself, beyond performing the assay according to established protocols. Its performance figures (correlation, regression, precision) are entirely standalone.

7. The Type of Ground Truth Used:

Reference Methods: The ground truth was established by two universally recognized and accepted reference methods for HDL-C measurement:
- The Phosphotungstic Acid MgCl (PTA) precipitation method
- The Center for Disease Control (CDC) reference method

8. The Sample Size for the Training Set:

Not Applicable (N/A) / No Distinct Training Set: This device is a chemical assay, not a machine learning algorithm. Therefore, there is no "training set" in the computational sense. The kit's reagents and protocol are developed through R&D and optimization, not by training on a dataset. The comparative and precision studies are for validation, not training.

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

Not Applicable (N/A): As there is no training set in the sense of machine learning, this question does not apply. The "ground truth" for developing the assay itself would come from biochemical principles, chemical validation, and performance optimization against known standards and reference methods during its development phase.

Summary

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ATTACHMENT 1

510(k) Summary Of Safety and Effectiveness Information Upon Which An Equivalence Determination Could be Made

Trade or Proprietary Name:	Genzyme N-geneousTM HDL Cholesterol Kit
Common or Usual Name:	Homogeneous assay for high density lipoprotein cholesterolCalibrator for high density lipoprotein cholesterol
Classification Name:	High density lipoprotein cholesterol testCalibrator, Primary
Manufacturer:	Genzyme DiagnosticsOne Kendall SquareCambridge, MA 02139-1562
Contact Person:	Nancy E. Isaac, Associate Director, Regulatory Affairs (617) 374-7431 orBeth A. Crowley, Regulatory Associate, (617) 252-7669.

The use of the Genzyme N-geneous™ HDL Cholesterol Kit in the clinical laboratory setting is substantially equivalent to the Phosphotungstic Acid MgCly (PTA) precipitation method.

The first readent contains a mixture of polymers and polvanions that bind to the sufface of low-density lipoproteins (LDL), very low-density lipoproteins (VLDL) and chylomicrons. These complexed lipoproteins are stabilized, even in the presence of detergent which is added as part of the second reagent, together with cholesterol enzymes. HDL particles, on the other hand, are not stabilized by the polymers and polvanions and become solubilized by the detergent. Consequently, only the HDL cholesterol is subject to cholesterol measurement.

Comparative performance studies were conducted at three sites using the N-geneous™ HDL Cholesterol Kit and two reference methods: Phosphotungstic Acid MgClo and the Center for Disease Control (CDC) reference method. Based on the Lipid Research Clinics (LRC) population studies, patient serum samples with HDL values between 27-91 mg/dL (5th and 95th percentively) were used for these comparative studies.

One hundred fourteen serum samples, with HDL values between 24 and 94 mg/dL, were tested at three sites with the N-geneous™ HDL test method. Of these 114 samples, 108 had sufficient volume

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for testing with the PTA predicate method. Additionally, a subset of at least 25 serum samples were Und i to be analyzed by the CDC reference method and the results compared.

	N-geneous™ vs.Phosphotungstic Acid(n = 108)	N-geneous™ vs.CDC Method(n = 26)
Correlation Coefficient (r)	0.96	0.96
Regression Equation	$y = 0.81x + 7.82$	$y = 1.01x - 3.39$

Precision studies were conducted using the N-geneous™ HDL Cholesterol Test Kit. Both within-run and between-run studies were performed using fresh and frozen serum pools at three target levels of HDL Cholesterol as determined by the NCEP: <35 mg/dL (low); 35-60 mg/dL (mid); and >60 mg/dL (high).

Within-Run	Low(<35 mg/dL)	Mid(35-60 mg/dL)	High(>60 mg/dL)
n	20	20	20
Mean (mg/dL)	30.2	42.7	75.6
SD (mg/dL)	0.45	0.50	0.79
%CV	1.50%	1.18%	1.04%

Between-Run	Low(<35 mg/dL)	Mid(35-60 mg/dL)	High(>60 mg/dL)
n	80	80	80
Mean (mg/dL)	29.4	41.9	73.2
SD (mg/dL)	1.11	1.10	2.03
%CV	3.79%	2.64%	2.78%

These data demonstrate that the performance of the N-geneous™ HDL Cholesterol Kit in the clinical laboratory is substantially equivalent to the performance of the PTA and CDC Reference methods.

In lieu of a 510(k) statement under 513(i) of the Act, this information is provided as a 510(k) summary for disclosure to any other persons/companies without the specific written authorization from Genzyme Corporation.

Regulation Number and Section

§ 862.1475 Lipoprotein test system.

(a)
Identification. A lipoprotein test system is a device intended to measure lipoprotein in serum and plasma. Lipoprotein measurements are used in the diagnosis and treatment of lipid disorders (such as diabetes mellitus), atherosclerosis, and various liver and renal diseases.(b)
Classification. Class I (general controls). The device is exempt from the premarket notification procedures in subpart E of part 807 of this chapter subject to § 862.9.