Reagents: For the direct, quantitative determination of low-density lipoprotein cholesterol (LDL-C) in human serum or plasma. Calibrator: For the calibration of the N-geneous® LDL-ST Cholesterol assay in serum or plasma. Controls: To monitor the performance of the Genzyme Direct LDL Cholesterol, N-geneous® LDL Cholesterol and N-geneous® LDL-ST Cholesterol Reagents.

Device Description

The Genzyme N-geneous® LDL-ST Cholesterol Reagent is a two-reagent homogeneous method for the direct quantitative determination of low density lipoprotein cholesterol (LDL-C) in human serum and plasma. This method is applicable to Olympus two-reagent Chemistry Analyzers and does not require any off-line pretreatment or centrifugation steps. The principle of the test is based upon a unique detergent which selectively solubilizes only the non-LDL lipoproteins, allowing them to be removed by cholesterol enzymes prior to the LDL cholesterol reaction. The LDL particles remain intact. The hydrogen peroxide produced by the reaction of the enzymes with the released cholesterol is consumed by a peroxidase reaction with 4-aminoantipyrine, yielding a colorless product. A second detergent, capable of releasing LDL cholesterol molecules, is added. The enzyme reaction with LDL cholesterol in the presence of the coupler produces color which is proportional to the amount of LDL cholesterol in the sample.

AI/ML Overview

The Genzyme N-geneous® LDL-ST Cholesterol Reagent is a two-reagent homogeneous method for the direct quantitative determination of low density lipoprotein cholesterol (LDL-C) in human serum and plasma. The study aimed to demonstrate the substantial equivalence of the Genzyme N-geneous® LDL-ST Cholesterol Reagent to the N-geneous® LDL Cholesterol Reagents and the ß-Quantification methods.

Acceptance Criteria and Reported Device Performance:

The document does not explicitly state pre-defined acceptance criteria values for the slope, intercept, or correlation coefficient. However, the study results are presented for comparison to the predicate devices. The implicit acceptance criteria are that the performance metrics of the N-geneous® LDL-ST on the Olympus AU600 are comparable to those of the predicate methods (ß-Quantification and N-geneous® LDL on Hitachi 911).

Metric	Acceptance Criteria (Implicit - based on predicate performance)	Reported Performance (N-geneous® LDL-ST vs. ß-Quantification)	Reported Performance (N-geneous® LDL-ST vs. N-geneous® LDL)
Slope	Expected to be close to 1	1.08	1.02
Intercept (mg/dL)	Expected to be close to 0	-8.7	-1.7
Correlation Coefficient (r)	Expected to be close to 1	0.969	0.990
Mean Difference (mg/dL)	Expected to be close to 0	0.6	0.3
Mean Percent Difference (%)	Expected to be close to 0	-0.1	0.1
Within-Run Precision %CV (Low)	Not explicitly stated, lower is better	1.4	N/A
Within-Run Precision %CV (High)	Not explicitly stated, lower is better	0.8	N/A
Between-Run Precision %CV (Low)	Not explicitly stated, lower is better	2.1	N/A
Between-Run Precision %CV (High)	Not explicitly stated, lower is better	2.7	N/A

Device Performance Study Details:

Sample Size and Data Provenance:
- Test Set for comparative studies:
  - 77 serum samples used for comparison between N-geneous® LDL-ST and N-geneous® LDL.
  - 62 of these 77 samples had sufficient volume for comparison with the ß-Quantification method.
- Test Set for precision studies: 20 replicates for each of the "Low" and "High" controls for both within-run and between-run precision.
- Data Provenance: The study was conducted at Genzyme Corporation. The country of origin of the samples is not specified, but the context implies a retrospective analysis of collected human serum samples.
Number of Experts and Qualifications: Not applicable. This is a quantitative chemical assay, not an imaging or diagnostic device requiring expert interpretation for ground truth.
Adjudication Method: Not applicable for a quantitative chemical assay.
Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study: Not applicable. This is a chemical assay, not a device requiring human reader interpretation in a clinical setting.
Standalone (Algorithm Only) Performance: The performance reported is the standalone performance of the Genzyme N-geneous® LDL-ST Cholesterol Reagent on the Olympus AU600 Analyzer. There is no human-in-the-loop component.
Type of Ground Truth Used:
- For the comparative studies, the ground truth was established by two reference laboratory methods:
  - ß-Quantification method (considered a gold standard for LDL-C measurement).
  - N-geneous® LDL Cholesterol Reagent (a previous version/predicate device from Genzyme).
- For the precision studies, the ground truth was based on the measured mean values of Genzyme Desirable and Risk controls.
Sample Size for Training Set: Not applicable. This is a chemical reagent, not a machine learning algorithm that requires a training set in the conventional sense. The development of the reagent itself would involve R&D and analytical optimization, but this document does not detail a "training set" for a predictive model.
How Ground Truth for Training Set was Established: Not applicable, as there is no specified training set for a machine learning model. The chemical formulation and methodology of the reagent are based on established biochemical principles and experimental validation during its development.

Summary

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K992766

510(k) PREMARKET NOTIFICATION

Genzyme Corporation
One Kendall Square
Cambridge, MA 02139

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

Trade or Proprietary Name:	Genzyme N-geneous® LDL-ST Cholesterol ReagentGenzyme N-geneous® LDL-ST Cholesterol CalibratorGenzyme LDL Control Set
Common or Usual Name:	Homogeneous assay for low density lipoprotein cholesterol
Classification Name:	Low density lipoprotein cholesterol testCalibrator, PrimaryLow density lipoprotein control
Manufacturer:	Genzyme DiagnosticsOne Kendall SquareCambridge, MA 02139-1562
Contact Person:	Robert Yocher, Vice President, Regulatory Affairs (617) 374-7275Barbara Pizza, Manager, Regulatory Affairs (617) 252-7953.

The use of the Genzyme N-geneous® LDL-ST Cholesterol Reagents in the clinical laboratory setting is substantially equivalent to the N-geneous® LDL Cholesterol Reagents and the B-Quantification methods.

The principle of the test is based upon a unique detergent which selectively solubilizes only the non-LDL lipoproteins, allowing them to be removed by cholesterol enzymes prior to the LDL cholesterol reaction. The LDL particles remain intact. The hydrogen peroxide produced by the reaction of the enzymes with the released cholesterol is consumed by a peroxidase reaction with 4-aminoantipyrine, yielding a colorless product. A second detergent, capable of releasing LDL cholesterol molecules, is added. The enzyme reaction with LDL cholesterol in the presence of the coupler produces color which is proportional to the amount of LDL cholesterol in the sample.

Comparative performance studies were conducted using the N-geneous® LDL-ST Cholesterol Reagents and two reference methods: N-geneous® LDL Cholesterol Reagent and ß-Quantification.

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510(k) PREMARKET NOTIFICATION

Seventy-seven serum samples, with LDL values between 54.5 and 222.1 mg/dL, were tested at Genzyme Corporation using the N-geneous® LDL-ST on the Olympus AU600 Analyzer and the Ngeneous® LDL methods. Of these 77 samples, 62 had sufficient volume to run ß-Quantification method.

N-geneous® LDL-ST on theOlympus AU600	vs.β-Quantification(n = 62)	vs.N-geneous® LDL onHitachi 911(n = 77)
Slope	1.08	1.02
Intercept (mg/dL)	-8.7	-1.7
Correlation Coefficient (r)	0.969	0.990
Mean (mg/dL)	LDL-ST: 119.6β-Q: 119.0	LDL-ST: 121.6LDL: 121.3
Standard Deviation (mg/dL)	LDL-ST: 28.9β-Q: 26.0	LDL-ST: 30.7LDL: 29.9
Mean Difference (mg/dL)	0.6	0.3
Mean Percent Difference (%)	-0.1	0.1

ß-Q = ß-Quantification

Precision studies were conducted using the N-geneous® LDL-ST Cholesterol Reagents on the Olympus Analyzers. Both within-run and between-run studies were performed using Genzyme Desirable and Risk controls:

Within-Run Precision	Low	High
	(<130 mg/dL)	(≥160 mg/dL)
n	20	20
Sample Range (mg/dL)	92.0 - 97.0	229.6 - 237.3
Mean (mg/dL)	93.8	233.5
SD (mg/dL)	1.3	1.9
%CV	1.4	0.8
Between run	Low(<130 mg/dL)	High(≥160 mg/dL)
n	20	20
Sample Range (mg/dL)	83 - 90	178 - 195
Mean (mg/dL)	86.9	188.4
SD (mg/dL)	1.8	5.0
%CV	2.1	2.7

These data demonstrate that the performance of the N-geneous® LDL-ST Cholesterol Reagents in the clinical laboratory is substantially equivalent to the performance of the N-geneous® LDL Cholesterol Reagent and the ß-Quantification methods.

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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.

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DEPARTMENT OF HEALTH & HUMAN SERVICES

Image /page/3/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a stylized eagle with three tail feathers, representing the three levels of government: federal, state, and local. The eagle is positioned to the right of the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA", which is arranged in a circular fashion around the eagle.

OCT 1 8 1999

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

Ms. Barbara Pizza Manager, Regulatory Affairs Genzyme Corporation One Kendall Square Cambridge, Massachusetts 02139-1562

Re: K992766

Trade Name: N-geneous® LDL-ST Cholesterol Reagent Regulatory Class: I reserved P Product Code: MRR N-geneous® LDL-ST Cholesterol Calibrator Regulatory Class: II Product Code: JIS Genzyme LDL Cholesterol Control Set Regulatory Class: I reserved Product Code: JJX Dated: August 16, 1999 Received: August 17, 1999

Dear Ms. Pizza:

We have reviewed your Section 510(k) notification of intent to market the device referenced above and we 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). 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.

If your device is classified (see above) into either class II (Special Controls) or class III (Premarket Approval), it may be subject to such additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 895. A substantially equivalent determination assumes compliance with the Current Good Manufacturing Practice requirements, as set forth in the Quality System Regulation (OS) for Medical Devices: General regulation (21 CFR Part 820) and that, through periodic QS inspections, the Food and Drug Administration (FDA) will verify such assumptions. Failure to comply with the GMP regulation may result in regulatory action. In addition, FDA may publish further announcements concerning your device in the Federal Register. Please note: this response to your premarket notification submission does not affect any obligation you might have under sections 531 through 542 of the Act for devices under the Electronic Product Radiation Control provisions, or other Federal laws or regulations.

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Page 2

Under the Clinical Laboratory Improvement Amendments of 1988 (CLIA-88), this device may require a CLIA complexity categorization. To determine if it does, you should contact the Centers for Disease Control and Prevention (CDC) at (770) 488-7655.

This letter will allow you to begin marketing your device as described in your 510(k) premarket notification. The FDA finding of substantial equivalence of your device to a legally marketed predicate device results in a classification for your device and thus, permits your device to proceed to the market.

If you desire specific advice for your device on our labeling regulation (21 CFR Part 801 and additionally 809.10 for in vitro diagnostic devices), please contact the Office of Compliance at (301) 594-4588. Additionally, for questions on the promotion and advertising of your device, please contact the Office of Compliance at (301) 594-4639. Also, please note the regulation entitled, "Misbranding by reference to premarket notification"(21 CFR 807.97). Other general information on your responsibilities under the Act may be obtained from the Division of Small Manufacturers Assistance at its toll-free number (800) 638-2041 or (301) 443-6597, or at its internet address "http://www.fda.gov/cdrlv/dsma/dsmamain.html".

Sincerely yours.

Steven Sutman

Steven I. Gutman, M.D, M.B.A. Director Division of Clinical Laboratory Devices Office of Device Evaluation Center for Devices and Radiological Health

Enclosure

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3.0 INTENDED USE

Page 1 of __ 1

510(k) Number (if known): K993746

Device Name: _N-geneous® LDL-ST Cholesterol Reagent

N-geneous® LDL-ST Cholesterol Calibrator

Genzyme LDL Cholesterol Control Set

Indications for Use:

Reagents:

For the direct, quantitative determination of low-density lipoprotein cholesterol (LDL-C) in human serum or plasma.

Calibrator:

For the calibration of the N-geneous® LDL-ST Cholesterol assay in serum or plasma.

Controls:

To monitor the performance of the Genzyme Direct LDL Cholesterol, N-geneous® LDL Cholesterol and N-geneous® LDL-ST Cholesterol Reagents.

Tom Coker
(Division Sign-Off)

Division of Clinical Laboratory Devices
510(k) Number K992761

RxV

CTC

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.