K Number

Device Name

VYSIS AUTOVYSION SYSTEM

Manufacturer

VYSIS

Date Cleared

2004-12-13

(61 days)

Product Code

NTH SYS

Regulation Number

866.4700

Intended Use

The Vysis® AutoVysion™ System is an automated scanning microscope and image analysis system. It is intended for in vitro diagnostic use with the Vysis® PathVysion® HER-2 DNA Probe Kit to aid in the detection and enumeration of FISH signals in interphase nuclei. and to determine the LSI® HER-2 to CEP® 17 signal ratio of the HER-2/neu gene via FISH in formalinfixed, paraffin-embedded human breast cancer tissue specimens. The AutoVysion System is intended to reduce overall hands-on time by performing automated enumeration. For a small percentage of samples (less than 7%) manual enumeration may be required. The Vysis® AutoVysion™ System is an adjunctive computer-assisted methodology to assist in the acquisition and measurement of images from microscope slides of formalin-fixed, paraffin-embedded breast cancer tissue sections for the presence of amplified HER-2/neu gene. The Vysis® AutoVysion™ System is intended for use as aid in determining HER-2/neu amplification status. in coniunction with optional manual visualization directly through the fluorescence microscope. When used with the Vysis® PathVysion® HER-2 DNA Probe Kit, the Vysis® AutoVysion™ System is indicated for use as a) an adjunct to existing clinical and pathologic information currently used as prognostic factors in stage II, node-positive breast cancer patients b) an aid to predict disease-free and overall survival in patients with stage II. node positive breast cancer treated with adjuvant cyclophosphamide, doxorubicin and 5-fluorouracil (CAF) chemotherapy; and, c) aid in the assessment of patients for whom HERCEPTIN® (Trastuzumab) treatment is being considered (see HERCEPTIN package insert).

Device Description

The Vysis® AutoVysion™ System consists of an automated fluorescence microscope with motorized scanning stage, a large-format monochrome CCD camera, computer and scanning and assay specific analysis software. The microscope is equipped with a mercury arc lamp for fluorescence epi-illumination; three single-pass fluorescence filter sets for DAPI, SpectrumGreen™ (SG) and SpectrumOrange™ (SO) and a triple-pass fluorescence filter set for DAPI/SG/SQ, all mounted in a motorized filter turret; 10x and 40x objectives in a motorized objective turret; 10x eyepieces; a CCD camera; and a motorized scanning stage that holds up to 8 slides. Images of single fluorescence colors are captured by the CCD camera and transferred to the computer. All functions are controlled by the System software.

More Information

Contact

Type

Center

Panel

Date Received

Product Code

Subsequent Product Codes

Applicant Name (Manufacturer)

Device Name

Decision

Street 1

Street 2

City

State

Country Code

ZIP

Postal Code

Class Advise Committee

SSP Indicator

Third Party Reviewed

Expedited Review

Predicate Devices

None

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The document mentions the use of the Expectation Maximization Algorithm (EM), which is a statistical algorithm, not typically classified as AI or ML in the context of modern medical device regulation. The description focuses on automated image analysis and enumeration, not learning or adaptive capabilities.

Therapeutic

No
The device is an in vitro diagnostic (IVD) system used to aid in the detection and enumeration of FISH signals in breast cancer tissue specimens and determine HER-2/neu amplification status, not to directly treat or diagnose a patient. It is explicitly for in vitro diagnostic use.

Diagnostic

Yes

The "Intended Use / Indications for Use" section explicitly states that the system is "intended for in vitro diagnostic use" and to "aid in the detection and enumeration of FISH signals... and to determine the LSI® HER-2 to CEP® 17 signal ratio... as aid in determining HER-2/neu amplification status." It also lists several diagnostic contexts for its use, such as aiding in prognostic factors and assessing patients for treatment.

SaMD (Software as a Medical Device)

The device description explicitly states that the system consists of hardware components including an automated fluorescence microscope, CCD camera, and computer, in addition to the software.

IVD (In Vitro Diagnostic)

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

Here's why:

Intended Use: The "Intended Use / Indications for Use" section explicitly states that the system is "intended for in vitro diagnostic use with the Vysis® PathVysion® HER-2 DNA Probe Kit".
Purpose: The system is used to aid in the detection and enumeration of FISH signals in human breast cancer tissue specimens to determine the HER-2/neu gene amplification status. This is a diagnostic process performed on biological samples outside of the body.
Clinical Context: The intended use also describes how the results are used in a clinical context, such as aiding in predicting survival and assessing patients for HERCEPTIN treatment.

All of these points align with the definition of an In Vitro Diagnostic device.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

The Vysis® AutoVysion™ System is an adjunctive computer-assisted methodology to assist in the acquisition and measurement of images from microscope slides of formalin-fixed, paraffin-embedded breast cancer tissue sections for the presence of amplified HER-2/neu gene. The Vysis® AutoVysion™ System is intended for use as aid in determining HER-2/neu amplification status. in coniunction with optional manual visualization directly through the fluorescence microscope.

When used with the Vysis® PathVysion® HER-2 DNA Probe Kit, the Vysis® AutoVysion™ System is indicated for use as

a) an adjunct to existing clinical and pathologic information currently used as prognostic factors in stage II, node-positive breast cancer patients
b) an aid to predict disease-free and overall survival in patients with stage II. node positive breast cancer treated with adjuvant cyclophosphamide, doxorubicin and 5-fluorouracil (CAF) chemotherapy; and,
c) aid in the assessment of patients for whom HERCEPTIN® (Trastuzumab) treatment is being considered (see HERCEPTIN package insert).

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

NTH

Device Description

Mentions image processing

Yes

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Fluorescence microscopy

Anatomical Site

Formalin-fixed, paraffin-embedded human breast cancer tissue specimens

Indicated Patient Age Range

Not Found

Intended User / Care Setting

Not Found

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)

Precision/Reproducibility:
The Vysis® AutoVysion™ System was evaluated for inter-site and day-to-day reproducibility at 3 clinical sites. The study consisted of a total of 36 specimen slides prepared from four human breast tissue specimens with varving levels of HER-2/neu gene amplification (one normal, one borderline, one moderate and one high amplification). Each site received three of each of the specimens randomized over three days. The optimal number of fields of view (FOV) was determined using 5, 7 and 10 fields of view. All three numbers of FOVs gave similar results. The ten FOVs were selected to be used for all precision studies. Day-to-day reproducibility showed no statistically significant differences in variance. Inter-site reproducibility showed no statistically significant differences.

Method comparison with predicate device:
Study type: Comparison to conventional manual microscopy.
Sample size: 234 clinical slides from 39 tumors with varying levels of HER-2/neu copy number.
Key results: Concordance was evaluated as the agreement between manually enumerated and the calculated HER-2 to CEP 17 signal ratio and the Vysis® AutoVysion™ System produced HER-2 to CEP 17 signal ratio. Among all tissue specimens with informative results for both methods, 92.5% (196/212) were correctly classified. Positive agreement was 96.0% and negative agreement was 89.2%. If samples with results in the equivocal range i.e. HER-2 to CEP 17 signal ratios between 1.5 and 3.0 were excluded from the calculation, total agreement was 98.8% (169/171) with 100% positive agreement and 97.5% negative agreement.
There were two false positive results by the AutoVysion™ System. When these two samples were repeated six times manually and by the scanner, both methods gave positive results in five of the six repeats.
The average bias for the manual enumeration ratio range of 1.18 to 4.49 was 0.472 (SD = 1.24), which was 11.7% of the average manual enumeration ratio of 4.05, meeting the acceptable error of ±15%.

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

Method comparison study:
Total agreement: 92.5% (196/212)
Positive agreement: 96.0%
Negative agreement: 89.2%
Total agreement (excluding equivocal range): 98.8% (169/171)
Positive agreement (excluding equivocal range): 100%
Negative agreement (excluding equivocal range): 97.5%

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.

None

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

§ 866.4700 Automated fluorescence

in situ hybridization (FISH) enumeration systems.(a)
Identification. An automated FISH enumeration system is a device that consists of an automated scanning microscope, image analysis system, and customized software applications for FISH assays. This device is intended for in vitro diagnostic use with FISH assays as an aid in the detection, counting and classification of cells based on recognition of cellular color, size, and shape, and in the detection and enumeration of FISH signals in interphase nuclei of formalin-fixed, paraffin-embedded human tissue specimens.(b)
Classification. Class II (special controls). The special control is FDA's guidance document entitled “Class II Special Controls Guidance Document: Automated Fluorescencein situ Hybridization (FISH) Enumeration Systems.” See § 866.1(e) for the availability of this guidance document.

Summary

510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY

A. 510(k) Number: K041875

B. Purpose for Submission: New device

C. Analyte:

Her2/neu gene copy number on formalin-fixed paraffin-embedded breast cancer specimens

D. Type of Test:

Computer-assisted image analyzer for fluorescence in situ hybridization (FISH)

E. Applicant:

Vysis, Inc.

F. Proprietary and Established Names: Vysis® AutoVysion™ System for PathVysion HER-2 DNA Kit

G. Regulatory Information:

1. Regulation section: 21 CFR 866.4700, Automated Fluorescent in situ Hybridization (FISH) Enumeration Systems
1. Classification: II
1. Product Code:

NTH, System, Automated Scanning Microscope and Image Analysis for fluorescence in situ hybridization (FISH) assays

1. Panel:
  Immunology 82

H. Intended Use:

1. Intended Use:
  The Vysis® AutoVysion™ System is an automated scanning microscope and image analysis system. It is intended for in vitro diagnostic use with the Vysis® PathVysion® HER-2 DNA Probe Kit to aid in the detection and enumeration of FISH signals in interphase nuclei. and to determine the LSI® HER-2 to CEP® 17 signal ratio of the HER-2/neu gene via FISH in formalinfixed, paraffin-embedded human breast cancer tissue specimens. The AutoVysion System is intended to reduce overall hands-on time by performing automated enumeration. For a small percentage of samples (less than 7%) manual enumeration may be required.

1. Indication(s) for use:

When used with the Vysis® PathVysion® HER-2 DNA Probe Kit, the Vysis® AutoVysion™ System is indicated for use as

a) an adjunct to existing clinical and pathologic information currently used as prognostic factors in stage II, node-positive breast cancer patients
b) an aid to predict disease-free and overall survival in patients with stage II. node positive breast cancer treated with adjuvant cyclophosphamide, doxorubicin and 5-fluorouracil (CAF) chemotherapy; and,
c) aid in the assessment of patients for whom HERCEPTIN® (Trastuzumab) treatment is being considered (see HERCEPTIN package insert).
1. Special condition for use statement(s): For prescription use.
1. Special instrument Requirements:
- Vysis® AutoVysion™ System

I. Device Description:

J. Substantial Equivalence Information:

1. Predicate device name(s) None
1. Predicate K number(s): None
1. Comparison with predicate: Not applicable

K. Standard/Guidance Document Referenced (if applicable):

FDA guidance documents on software validation and off-the shelf Software use and NCCLS- EP9-A2

L. Test Principle:

A qualified user visually inspects the tumor regions of the slide, previously identified by a pathologist, identifies areas of tumor invasion with acceptable hybridization quality and records the coordinates of those areas for analysis through a point and click interface. Once the target areas have been identified, the AutoVysion™ System enters a fully automatic process, capturing extended-focus images of the marked areas at 40x magnification in each color: DAPI. SpectrumGreen and SpectrumOrange. All image data is saved to disk. The hybridization signals in each area are detected and enumerated automatically. The slide is also assessed for appropriate hybridization quality requirements that, if not satisfied, the sample may be rejected for automatic

analysis. Final review and reporting of sample results is performed by a qualified user.

The system uses a "targeted tiles" method for sampling the tumor. In this method, each field of view (FOV) in the area selected for analysis is sampled by placing a set of non-overlapping square "tiles" of equal size on the image. Each tile is comparable in size to the area of a tumor cell nucleus. The tiles are placed one by one in a way that maximizes the DAPI fluorescence contained in each tile, so that the set of tiles covers much of the nuclear material in the FOV. The spot count of a particular tile comprises the total spot count of the cell nuclei that are wholly or partly incorporated in the tile randomly reduced by truncation by tile boundary and microtome slicing. The method used to analyze the observed distribution of per-tile spot counts is the Expectation Maximization Algorithm (EM). EM is used to fit a mixture of two distributions to the observed two dimensional spot count distribution. The goodness of fit of the two-distribution model is compared with the goodness of fit of a single distribution to ensure that truly homogeneous samples are not erroneously fitted by two separate distributions. The HER-2/CEP 17 ratio is obtained directly from the parameters of the fitted distribution(s).

M. Performance Characteristics (if/when applicable):

1. Analytical performance:
- a. Precision/Reproducibility:

The Vysis® AutoVysion™ System was evaluated for inter-site and day-to-day reproducibility at 3 clinical sites. The study consisted of a total of 36 specimen slides prepared from four human breast tissue specimens with varving levels of HER-2/neu gene amplification (one normal, one borderline, one moderate and one high amplification). Each site received three of each of the specimens randomized over three days. The optimal number of fields of view (FOV) was determined using 5, 7 and 10 fields of view. All three numbers of FOVs gave similar results. The ten FOVs were selected to be used for all precision studies.

Day-to-day reproducibility was determined by calculating the mean observed ratio of LSI HER-2/neu to CEP 17. standard deviation (SD) and percent coefficient of variation (%CV) generated from 10 fields of view for each specimen across the three study days. The p-values associated with the Levene test statistics were calculated to test the homogeneity of day-to-day variances, with a 0.05 significance level. Results showed no statistically significant differences (see table below).

| Expected | Observed ratios of LSI HER-2/
neu to CEP 17 | | | P-value | | | | | | |
|----------|------------------------------------------------|------|-------|---------|------|-------|-------|------|-------|--------|
| | Day 1 | | | Day 2 | | | Day 3 | | | |
| | Mean | SD | %CV | Mean | SD | %CV | Mean | SD | %CV | |
| 1.33 | 1.17 | 0.33 | 28.04 | 1.11 | 0.14 | 12.3 | 1.14 | 0.06 | 5.12 | 0.1152 |
| 1.71 | 2.08 | 0.61 | 29.34 | 2.38 | 0.64 | 26.82 | 2.45 | NR | NR | 0.9049 |
| 8.14 | 5.70 | 0.86 | 15.01 | 5.55 | 0.33 | 5.87 | 5.47 | 0.66 | 12.09 | 0.2788 |
| 12.97 | 6.42 | 0.81 | 12.55 | 7.42 | 0.17 | 2.24 | 8.01 | 0.35 | 4.38 | 0.1205 |

NR= No result

Inter-site reproducibility was similarly determined across the three study sites. Results for the three sites were not statistically significant and are summarized in the following table:

Expected	Observed ratios of LSI HER-2/neu to CEP 17			P-value

	Site 1			Site 2			Site 3
	Mean	SD	%CV	Mean	SD	%CV	Mean	SD	%CV
1.33	1.24	0.26	21.09	1.05	0.05	4.52	1.14	0.18	15.97	0.1833
1.71	2.09	0.58	27.84	2.42	NR	NR	2.39	0.75	31.23	0.5436
8.14	5.41	0.58	10.70	5.88	0.06	1.02	5.44	0.88	16.12	0.1612
12.97	6.95	1.32	19.03	7.51	0.81	10.72	7.39	0.30	4.04	0.1665

NR= No result

b. Linearity/assay reportable range: Not applicable.
Traceability (controls, calibrators, or method): C. The analytical traceability of the system depends on the Vysis® PathVysion® HER-2 DNA Probe Kit. The AutoVysion™ System employs ProbeCheck control slides for every run to assess the accuracy of signal enumeration and to monitor the assay performance.
d. Detection limit (functional sensitivity): Not applicable
e. Analytical specificity The specificity of the test result is dependent on the analytical performance of the Vysis® PathVysion® HER-2 DNA Probe Kit
f. Assay cut-off:

The assay cut-off of the test result is dependent on the analytical performance of the Vysis® PathVysion® HER-2 DNA Probe Kit.

1. Comparison studies:
- Method comparison with predicate device: a.

The substantial equivalence studies were based on comparison to conventional manual microscopy performed in accordance with Vysis® PathVysion® HER-2 DNA Probe Kit.

Duplicate slides from each tumor were randomized and assayed with the Vysis® PathVysion® HER-2 DNA Probe Kit according to the package insert instructions prior to shipment to the study sites. The randomized slides were enumerated by the standard and test method at each of the three study sites. Two hundred thirty-four clinical slides from 39 tumors with varying levels of HER-2/neu copy number were used in the study.

Concordance was evaluated as the agreement between manually enumerated and the calculated HER-2 to CEP 17 signal ratio and the Vysis® AutoVysion™ System produced HER-2 to CEP 17 signal ratio. Among all tissue specimens with informative results for both

methods, 92.5% (196/212) were correctly classified. Positive agreement was 96.0% and negative agreement was 89.2%. If samples with results in the equivocal range i.e. HER-2 to CEP 17 signal ratios between 1.5 and 3.0 were excluded from the calculation, total agreement was 98.8% (169/171) with 100% positive agreement and 97.5% negative agreement.

	Manual
Scanner	1.5	1.5-10.0
------------	---------	----------	-------
Mean	3.35	7.39	12.77
SD	0.95	1.41	1.80
N	33	42	16

N. Instrument Name:

Vysis® AutoVysion™ System

O. System Descriptions:

See (H) Device Description.

Modes of Operation: 1.
Semi-automated computer-assisted interpretation.
1. Software:
  FDA has reviewed applicant's Hazard Analysis and software development processes for this line of product types: Yes
1. Sample Identification: Slide identification is entered manually into the AutoVysion™ System before the slides are loaded into the instrument.
1. Specimen Sampling and Handling:

The microscope slides to be examined are loaded onto the microscope stage of the AutoVysion™ System and the user records the coordinates of those areas for analysis through a point and click interface. Once the target areas have been identified, the AutoVysion™ System automatically captures images of the marked areas in each fluorescence color and enumerates the hybridization

signals in each area. The system also rejects slide that failed hybridization quality requirements for automatic analysis.

న్. Assay Types: Computer-assisted image analysis of fluorescence in situ hybridization signals in interphase nuclei of cells in formalin-fixed paraffin-embedded tissue.
1. Reaction Types: Fluorescent microscopy
1. Calibration:

The AutoVysion™ instrument is factory calibrated. Monthly calibration checks with End Switches and Movements tests should be performed. To assess accuracy of signal enumeration by the instrument, laboratory-stained Vysis ProCheck slides are used for every staining run.

1. Quality Control:
  The accuracy of the system depends on the laboratory following the quality control instructions recommended in the labeling of the fluorescence in situ hybridization (FISH) assay kit associated with the AutoVysion™.

P. Other Supportive Instrument Performance Characteristics Data Not Covered In The "M. Performance Characteristics" Section Of The SE Determination Decision Summary.

None. Q. Conclusion:

The petition for Evaluation of Automatic Class III Designation for this device is accepted. The device is classified as Class II under regulation 21 CFR 866.4700 with special controls. The special control guidance document " Class II Special Controls Guidance Document: Automated Fluorescence in situ Hvbridization (FISH) Enumeration Systems" is available at WWW.fda.gov/cdrh/oivd/guidance/1550.pdf.