ICONOSCOPE ONCOFISH HER2 TEST SYSTEM, MODEL 2000

K080909 · Ikonisys, Inc. · NTH · Oct 17, 2008 · Immunology

Device Facts

• Record ID: K080909
• Device Name: ICONOSCOPE ONCOFISH HER2 TEST SYSTEM, MODEL 2000
• Applicant: Ikonisys, Inc.
• Product Code: NTH · Immunology
• Decision Date: Oct 17, 2008
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.4700
• Device Class: Class 2

Indications for Use

The Ikoniscope® oncoFISH® her2 Test System is an automated scanning microscope coupled with image analysis, acquisition, and display functions. It is intended for in-vitro diagnosis as an aid to the technologist or pathologist in the detection, classification, and enumeration of cells of interest based on particular characteristics such as intensity, size, shape, or fluorescence. The Ikoniscope® oncoFISH® her2 Test System is intended to detect and quantify chromosome 17 and the HER-2 gene via fluorescence in situ hybridization (FISH) in interphase nuclei from formalin-fixed, paraffin embedded human breast cancer tissue specimens, probed with the Abbott PathVysion® HER-2 DNA Probe Kit. The Ikoniscope® oncoFISH® her2 Test System is to be used as an adjunctive automated enumeration tool, in conjunction with manual visualization, to assist in determining Her-2 gene to chromosome 17 signal ratio.

Device Story

The Ikoniscope® oncoFISH® her2 Test System is an automated scanning microscope for FISH signal enumeration in breast cancer tissue. The system accepts slides prepared with PathVysion® HER-2 DNA probes, loaded into a cassette. It performs automated slide loading, scanning at multiple magnifications (4X, 20X, 100X), and digital image acquisition. An automated algorithm detects and enumerates fluorescent signals for HER-2 and CEP 17 genes. The system calculates the HER-2/CEP 17 ratio and presents an image gallery of cells to the operator. The pathologist or technologist reviews the automated results to make a diagnostic determination of HER-2 amplification status. The device is intended for clinical laboratory use to assist in breast cancer diagnostics.

Clinical Evidence

Performance evaluated via method comparison and reproducibility studies. Method comparison (n=182 slides) against manual PathVysion read showed 94.5% overall agreement, 91.5% positive percent agreement, and 97% negative percent agreement. Reproducibility study across 3 clinical sites using 6 breast cancer specimens showed site-to-site concordance of 83.3%. Scan-to-scan variation was assessed with %CV ranging from 3.06% to 11.49%.

Technological Characteristics

Automated scanning microscope system; epi-fluorescent microscopy; digital image capture; magnification 4X, 20X, 100X; barcode specimen identification; automated slide loading/handling; software-based signal enumeration; moderate level of concern software.

Indications for Use

Indicated for use by technologists or pathologists as an adjunctive tool for the detection, classification, and enumeration of HER-2 gene and chromosome 17 signals in interphase nuclei from formalin-fixed, paraffin-embedded human breast cancer tissue specimens. Used with Abbott PathVysion® HER-2 DNA Probe Kit. Not for breast cancer screening or diagnosis.

Regulatory Classification

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.

Special Controls

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 Hybridization (FISH) Enumeration Systems" is available at www.fda.gov/cdrh/oivd/guidance/1550.pdf.

*Classification.* Class II (special controls). The special control is FDA's guidance document entitled “Class II Special Controls Guidance Document: Automated Fluorescence*in situ* Hybridization (FISH) Enumeration Systems.” See § 866.1(e) for the availability of this guidance document.

Predicate Devices

• Bioview Duet™ (k050840, k061602)
• Vysis® AutoVysion™ System (K041875)

Submission Summary (Full Text)

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# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY

A. 510(k) Number:
k080909

B. Purpose for Submission:
New device

C. Manufacturer and Instrument Name:
Ikonisys, Inc., Ikoniscope® oncoFISH™ her2 Test System

D. Type of Test or Tests Performed:
An automated system for enumeration of fluorescence in situ hybridization (FISH) signals of HER-2/neu gene in human breast cancer specimens.

E. System Descriptions:

1. Device Description:
The Ikoniscope® oncoFISH her2 Test System is an automated scanning microscope system incorporating automated slide loading and handling, low and high magnification scanning to identify targets of interest and digital image acquisition, coupled with an image analysis workstation. Microscope slides, prepared according to the DNA probe manufacturers' specifications, are placed into a multiple slide cassette, and loaded into the Ikoniscope® oncoFISH her2 Test system. The system unloads each slide, scans each one, and returns it to the cassette automatically. During scanning, images of cells exhibiting the predetermined characteristics for FISH signals are digitally photographed and stored. After all the slides are scanned, the workstation provides an image gallery for each slide that displays the image of each cell meeting predetermined characteristics and quantity and places scanned nuclei into scorable categories, established according to the specifications in the DNA probes FDA cleared labeling. The operator/reader can then evaluate the cell nuclei, and make the diagnostic determination accordingly.

2. Principles of Operation:
The Ikoniscope® oncoFISH her2 Test System combines elements of existing technologies to perform its function. Fluorescence In-Situ Hybridization (FISH) uses commercially available, FDA cleared, DNA probes (not supplied with the test system). Samples are prepared according to the instructions for the Vysis® PathVysion™ HER-2 DNA Probe kit. The user selects the appropriate areas for analysis in accordance with the PathVysion™ kit instructions. The oncoFISH system scans and captures images for each of the selected areas and the automatic algorithm detects and enumerates the fluorescent signals. The user reviews the signal enumeration for all relevant cells and the results (total number of cells and overall signal ratio) are automatically calculated by the system.

There is no change in the system hardware from the previously cleared system. Software changes were implemented to support the new indication for use for HER-2/neu FISH automated enumeration. The device methodology is well

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established.

3. Modes of Operation:

Semi-automated

4. Specimen Identification:

Barcode

5. Specimen Sampling and Handling:

Samples should be obtained and handled according to the laboratory's standard operating procedures and following the protocol described in the package insert for the PathVysion probe kit.

6. Calibration:

Calibration of the Ikoniscope is done at the time of installation by Ikonysis.

7. Quality Control:

ProbeChek® quality control slides by Abbott should be used with the PathVysion probes as recommended by the probe manufacturer.

8. Software:

FDA has reviewed applicant's Hazard Analysis and Software Development processes for this line of product types:

Yes ☐ X ☐ or No ☐ Comprehensive software documentation at a

Moderate Level of Concern was provided

F. Regulatory Information:

| Product Code | Classification | Regulation Section | Panel |
| --- | --- | --- | --- |
| NTH | Class II | 21 CFR 866.4700 Automated fluorescent in situ hybridization (FISH) enumeration system | Immunology 82 |

G. Intended Use:

1. Indication(s) for Use:

The Ikoniscope® oncoFISH® her2 Test System is an automated scanning microscope coupled with image analysis, acquisition and display functions. It is intended for in-vitro diagnosis as an aid to the technologist or pathologist in the detection, classification, and enumeration of cells of interest based on the ratio of HER-2 genes to CEP 17 genes. The Ikoniscope® oncoFISH® her2 Test System is intended to detect amplification of the HER-2/neu breast gene via fluorescence in situ hybridization (FISH) in formalin-fixed, paraffin-embedded human breast cancer tissue specimens.

2. Special Conditions for Use Statement(s):

For use only with PathVysion® HER-2 DNA Probe Kit (Abbott Molecular, Inc., Des Plaines, IL)

H. Substantial Equivalence Information:

1. Predicate Device Name(s) and 510(k) numbers:

BioView Duet™ System k050840, k061602

2. Comparison with Predicate Device:

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| Similarities | | |
| --- | --- | --- |
| Item | Device | Predicate |
| Intended Use | The Ikoniscope® oncoFISH® her2 Test System is an automated scanning microscope coupled with image analysis, acquisition and display functions. It is intended for in-vitro diagnosis as an aid to the technologist or pathologist in the detection, classification, and enumeration of cells of interest based on the ratio of HER-2 genes to CEP 17 genes. The Ikoniscope® oncoFISH® her2 Test System is intended to detect amplification of the HER-2/neu Breast gene via fluorescence in situ hybridization (FISH) in formalin-fixed, paraffin-embedded human breast cancer tissue specimens. | Same |
| Staining | Fluorescence In Situ Hybridization (FISH) | Same |
| Method of operation | Automated epi-fluorescent microscopy digital image capture of wave-length specific signals fluorescent signals | Same |
| Specimen Presentation | Automated | Same |
| Specimen Processing | Automated | Same |
| Clinical Decision Point | Amplified/Non-amplified | Same |
| Specimen Type | Tissue | Same |
| Enumeration | Ratio | Same |
| Differences | | |
| --- | --- | --- |
| Item | Device | Predicate |
| Magnification | Microscope: 4X, 20X, 100X | Microcopy: 10X, 40X |

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I. Special Control/Guidance Document Referenced (if applicable):

Determination of Intended Use for 510(k) Devices; Guidance for CDRH Staff
Guidance for Content of Premarket Submissions of Software Contained in Medical Devices FDA - General Principles of Software Validation
Medical Device Requirements - Human Factors and the FDA
FDA Guidance Document for In Vitro Diagnostics Devices that utilizes In Situ Hybridization technology for detection of Somatic Mutations – 1996
FDA Good Automated Manufacturing Practices – Guide for Validation of Automated Systems (ISPE/GAMP Forum, 2001)
FDA Guidance Document for Off-The Shelf Software Use in Medical Devices - 1999

J. Performance Characteristics:

1. Analytical Performance:

a. Accuracy:

Four clinical laboratories provided previously manually enumerated PathVysion slides for the method comparison study. The PathVysion slides from the four collection sites were tested at two (2) clinical laboratories with two (2) Ikoniscope® oncoFISH her2 systems. Each collection site provided 25 consecutive amplified (positive) and 25 non-amplified (negative) PathVysion slides for oncoFISH her2 testing. The H&E slides with the scribed area of interest for each corresponding PathVysion slide were used for the oncoFISH her2 testing. A total of 182 slides were enumerated by the PathVysion manual and oncoFISH her2 methods. The slides consisted of 100 non-amplified slides with a HER2/CEP17 ratio of 0.8 – 1.91 and 82 amplified slides with a HER2/CEP17 ratio of 2.02 – 11.3 by manual PathVysion. Reasons for the discordant results were poor slide quality, high background or few cells (n=3); operator not properly selecting amplified images from scan (n=3) or region or interest (ROI) identified on the H&E and PathVysion slides differ (n=1).

Overall Agreement was 94.5% (172/182), Positive Percent Agreement 91.5% (75/82) and Negative Percent Agreement 97% (97/100).

Table 1. Ikoniscope oncoFISH her2 vs. PathVysion Manual Read

| | Manual PathVysion | | |
| --- | --- | --- | --- |
| | Amplified | Non-amplified | Total |
| Ikoniscope oncoFISH her2 | Amplified | 75 | 3 |
| Non-amplified | 7 | 97 | 104 |
| Total | 82 | 100 | 182 |

b. Precision/Reproducibility:

A panel of six (6) breast cancer specimens, consisting of 2 each with HER2/CEP17 ratios of 0.8 – 1.7; 1.8 – 2.2; and > 2.2 by previously performed manual PathVysion were evaluated at 3 clinical sites over 3 non-consecutive

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days. Manual PathVysion results were determined after which the samples were evaluated with the oncoFISH her2 system. Six slides could not be evaluated due to poor quality. Results of the reproducibility study are shown below in Table 2.

Table 2. Reproducibility Results.

| | | Manual PathVysion HER2/CEP17 ratio | | | Ikoniscope oncoFISH HER2/CEP17 ratio | | |
| --- | --- | --- | --- | --- | --- | --- | --- |
| Specimen | Statistic | Site 1 | Site 2 | Site 3 | Site 1 | Site 2 | Site 3 |
| 1 | Mean | 1.83 | 1.51 | 1.83 | 1.9 | 1.48 | 1.92 |
| 1 | SD | 0.15 | 0.12 | 0.01 | 0.05 | 1.013 | 0.34 |
| 1 | C.V. (%) | 8.33 | 8.03 | 0.31 | 2.41 | 8.63 | 17.68 |
| 1 | N | 3 | 3 | 3 | 3 | 3 | 3 |
| 2 | Mean | 1.77 | 1.14 | 1.69 | 1.98 | 1.23 | 1.5 |
| 2 | SD | 0.06 | 0.1 | 0.06 | 0.15 | 0.13 | 0.06 |
| 2 | C.V. (%) | 3.27 | 8.68 | 3.29 | 7.52 | 10.35 | 3.92 |
| 2 | N | 3 | 3 | 3 | 2 | 2 | 3 |
| 3 | Mean | 1.17 | 0.99 | 1.12 | 1.16 | 1 | 1.54 |
| 3 | SD | 0.06 | 0.01 | 0.1 | 0.06 | 0.05 | 0.26 |
| 3 | C.V. (%) | 4.95 | 1.17 | 9.32 | 4.8 | 4.49 | 16.92 |
| 3 | N | 3 | 3 | 3 | 3 | 3 | 3 |
| 4 | Mean | 1.2 | 0.9 | 1.13 | 1.36 | 0.83 | 1.36 |
| 4 | SD | 0.1 | 0.02 | 0.07 | 0.03 | 0.1 | 0.03 |
| 4 | C.V. (%) | 8.33 | 2.37 | 6.26 | 2.55 | 11.93 | 1.85 |
| 4 | N | 3 | 2 | 2 | 3 | 2 | 3 |
| 5 | Mean | 10.47 | 6.01 | 7.75 | 11.92 | 6.67 | 10.47 |
| 5 | SD | 1.84 | 0.31 | 1.83 | 0.85 | 0.69 | 0.77 |
| 5 | C.V. (%) | 17.63 | 5.08 | 23.68 | 7.1 | 10.38 | 7.31 |
| 5 | N | 3 | 3 | 3 | 3 | 3 | 3 |
| 6 | Mean | 2.1 | 1.33 | 2.13 | 1.75 | 1.07 | 1.56 |
| 6 | SD | 0.1 | 0.26 | 0.43 | 0.25 | 0.01 | 0.31 |
| 6 | C.V. (%) | 4.76 | 19.27 | 20.15 | 14.26 | 0.66 | 19.64 |
| 6 | N | 3 | 3 | 3 | 3 | 2 | 3 |

Sites 1 and 3 each had discordant results between manual PathVysion and oncoFISH her2 for slide #6, while Site 2 reported this slide as non-amplified for both manual PathVysion and oncoFISH. Slides 1 and 2 were indeterminate (HER2/CEP17 = 1.8 - 2.2) based on their original PathVysion results but in this reproducibility study, these two slides were found non-amplified by both manual PathVysion (mean ratios 1.14 - 1.83) and oncoFISH (mean ratios = 1.23 - 1.98) by all three sites. Two (2) manual PathVysion slides were not readable due to no cells to score or failed hybridization. Four (4) Ikoniscope oncoFISH slides could not be analyzed with one had no cells to score, one with insufficient cells and two

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unacceptable scan. Based on final results, concordance between sites was $83.3\%$ .

Scan-to-scan variation with the Ikoniscope® oncoFISH her2 system was performed at one (1) clinical site. One daily panel of slides was re-scanned two (2) additional times on different days with the Ikoniscope yielding results from three (3) Ikoniscope® oncoFISH her2 runs. The table below (Table 3) lists the oncoFISH her2 ratio results with the mean, standard deviation (SD) and percent coefficient of variation (%CV) for the three scans of the same six (6) specimens.

Table 3. Scan-to-Scan Variation Results

| Specimen | N | Ikoniscope oncoFISH her2 HER2/CEP17 ratio | | |
| --- | --- | --- | --- | --- |
| | | Mean | SD | %CV |
| 1 | 3 | 1.63 | 0.05 | 3.06 |
| 2 | 3 | 1.63 | 0.07 | 4.79 |
| 3 | 3 | 1.31 | 0.08 | 6.54 |
| 4 | 3 | 1.45 | 0.167 | 11.49 |
| 5 | 3 | 10.36 | 0.56 | 5.49 |
| 6 | 3 | 1.66 | 0.1 | 6 |

c. Linearity: Not Applicable
d. Carryover: Not Applicable
e. Interfering Substances: Not Applicable

# K. Proposed Labeling:

The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10.

# L. Conclusion:

The submitted information in this premarket notification is complete and supports a substantial equivalence decision.