The Virtuoso TM system provides automated digital slide creation, management, analysis, and viewing. It is intended for in vitro diagnostic use as an aid to the pathologist in the display, detection, counting, review and classification of tissues and cells of clinical interest based on particular morphology, color, intensity, size, pattern and shape.

The Virtuoso™ System for IHC PR (1E2) using the VENTANA iScan HT is for the digital read application. This particular Virtuoso system is intended for use as an aid to the pathologist in the qualitative detection of progesterone receptor (PR) protein in formalin-fixed, paraffin-embedded normal and neoplastic tissue. This device is an accessory to Ventana Medical Systems, Inc. CONFIRM anti-Progesterone Receptor (PR) (1E2) Rabbit Monoclonal Primary Antibody assay. The CONFIRM™ anti-Progesterone Receptor (PR) (1E2) Rabbit Monoclonal Primary Antibody assay is indicated for use as an aid in the assessment of breast cancer patients for whom endocrine treatment is being considered (but is not the sole basis for treatment).

Note: The IHC PR (1E2) Digital Read application is an adjunctive computer-assisted methodology for the qualified pathologist in the acquisition and interpretation of images from microscope glass slides of breast cancer specimens stained for the presence of PR protein. The accuracy of the test results depends on the quality of the immunohistochemical staining. It is the responsibility of a qualified pathologist to employ appropriate morphological studies and controls as specified in the instructions for the CONFIRM™ anti-Progesterone Receptor (PR) (1E2) Rabbit Monoclonal Primary Antibody used to assure the validity of the Virtuoso System for IHC PR Digital Read scores. The actual correlation of CONFIRM™ anti-PR antibody to clinical outcome has not been established. This device is intended for IHC slides stained on the BenchMark ULTRA stainers. For prescription use only.

Device Description

The Virtuoso™ System is an instrument-plus-software system designed to assist the qualified pathologist in the consistent assessment of protein expression in immunohistochemically stained histologic sections from formalin-fixed, paraffinembedded normal and neoplastic tissues. The system consists of a slide scanner, computer, monitor, keyboard, and mouse for specific immunohistochemical markers, and software with a Windows web browser-based user interface. Virtuoso is a web-based, end-to-end, digital pathology software solution that allows pathology laboratories to acquire, manage, view, analyze, share, and report digital images of pathology specimens. Using the Virtuoso software, the pathologist can view digital images, add annotations and generate reports.

Hardware: The iScan HT scanning device captures digital images of formalinfixed, paraffin-embedded tissues that are suitable for storage and viewing. The device includes a digital slide scanner, a carousel for loading glass slides, computer, scanner software, keyboard, mouse and monitor.

Software: The Virtuoso software is designed to complement the routine workflow of a qualified pathologist in the review of immunohistochemically stained histologic slides. The software makes no independent interpretations of the data and requires competent human intervention for all steps in the process.

AI/ML Overview

Acceptance Criteria and Device Performance for Virtuoso™ System for IHC PR (1E2) using the VENTANA iScan HT

This response summarizes the acceptance criteria and study findings for the Virtuoso™ System for IHC PR (1E2) using the VENTANA iScan HT, based on the provided FDA 510(k) summary (K142965).

1. Table of Acceptance Criteria and Reported Device Performance

The acceptance criteria are implicitly derived from the reported agreement rates. For the purpose of this table, we will highlight the reported agreement rates as the measure of meeting the acceptance of substantial equivalence.

Performance Metric	Acceptance Criteria (Implicit from Predicate Equivalence)	Reported Device Performance (95% CI)	Study Type
Digital Read (DR) vs Manual Read (MR) Agreement (Overall Percent Agreement - OPA)	High agreement with manual microscopic evaluation (established by predicate devices)	Site 1: 98.7% (95.5-99.7)	Method Comparison
		Site 2: 88.9% (83.1-92.9)
		Site 3: 96.2% (91.9-98.2)
DR vs MR Agreement (Positive Percent Agreement - PPA)	High agreement with manual microscopic evaluation	Site 1: 99.1% (94.9-99.8)	Method Comparison
		Site 2: 86.7% (77.8-92.4)
		Site 3: 91.9% (83.4-96.2)
DR vs MR Agreement (Negative Percent Agreement - NPA)	High agreement with manual microscopic evaluation	Site 1: 98.0% (89.7-99.7)	Method Comparison
		Site 2: 91.1% (82.8-95.6)
		Site 3: 100.0% (95.5-100.0)
Intra-Pathologist/Inter-Day Reproducibility (DR - OPA)	High agreement across multiple reads by the same pathologist	Read 1 vs Read 2: 97.4% (86.8-99.5)	Reproducibility
		Read 1 vs Read 3: 89.7% (76.4-95.9)
		Read 2 vs Read 3: 92.3% (79.7-97.3)
Inter-Pathologist Reproducibility (DR - OPA)	High agreement between different pathologists using DR	Site 1 vs Site 2: 81.1% (74.3-86.5)	Reproducibility
		Site 1 vs Site 3: 77.4% (70.2-83.3)
		Site 2 vs Site 3: 89.9% (84.2-93.7)
Inter-Scanner Precision (OPA)	High agreement between different scanners	Site 1 vs Site 2: 90.0% (86.5-92.7)	Scanner Precision
		Site 1 vs Site 3: 93.6% (90.6-95.7)
		Site 2 vs Site 3: 90.3% (86.8-92.9)
Intra-Scanner/Inter-Day Precision (OPA)	High agreement on the same scanner across different days	Day 1 vs Day 2: 92.2% (89.0-94.6)	Scanner Precision
		Day 1 vs Day 3: 90.8% (87.4-93.4)
		Day 2 vs Day 3: 90.8% (87.4-93.4)

Note: The document states that the test system was shown to be "as safe and effective (therefore substantially equivalent) as the predicate devices". The provided agreement rates are the key metrics demonstrating this substantial equivalence. Specific predefined numerical acceptance criteria are not explicitly stated as distinct thresholds in the provided text but are implicitly met by achieving high concordance with manual methods and good reproducibility, consistent with existing legally marketed devices.

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

Sample Size for Method Comparison Test Set:
- Site 1: 159 cases
- Site 2: 162 cases
- Site 3: 156 cases
- Overall: 477 cases (sum of evaluable cases from the three sites)
Sample Size for Reproducibility Test Set: 39 cases (for intra-pathologist reproducibility)
Sample Size for Scanner Precision Test Set: 40 cases
Data Provenance: The document does not explicitly state the country of origin. However, the study involved three different "sites", implying a multi-center study possibly within the US, but this is not explicitly confirmed. The study appears to be retrospective in nature, using existing formalin-fixed, paraffin-embedded tissue blocks that were then stained and digitally scanned.

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

Number of Experts: 3 pathologists (referred to as "Readers" or "Investigators"). The method comparison study involved each of these 3 pathologists providing both manual and digital reads. The reproducibility studies also involved these 3 pathologists.
Qualifications of Experts: The document refers to them as "qualified pathologists" or "investigators." Specific details regarding their years of experience or sub-specialty (e.g., radiologist, breast pathologist) are not provided.

4. Adjudication Method for the Test Set

Method Comparison: The ground truth for the device's performance was established using the manual read (MR) by the same pathologist as the reference for the digital read (DR) evaluation. Each pathologist's DR results were compared to their own MR results. There is no explicit mention of an independent adjudication committee or consensus among multiple experts for the ground truth itself.
Reproducibility: For intra-pathologist reproducibility, the comparison was between the same pathologist across three different reading sessions. For inter-pathologist reproducibility, the comparison was between pairs of pathologists.
Scanner Precision: The comparison was between clinical scoring categories agreed upon at different sites/days for the same FOVs.
It appears no formal "N+1" or similar adjudication method was employed to establish a single, definitive ground truth independent of the readers whose digital reads were being assessed. Instead, the agreement between the digital read and a human's manual read served as the primary performance indicator, and reproducibility between humans (both manual and digital) was also evaluated.

5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study Was Done, and Effect Size

Yes, a form of MRMC study was implicitly done. The method comparison study involved 3 pathologists (multi-reader) evaluating multiple cases (multi-case), where their digital reads were compared to their manual reads. The reproducibility studies also involved multiple readers and multiple cases.
Effect Size (Human Reader Improvement with AI vs. without AI): The document does not report an effect size for human readers improving with AI assistance. The study design is primarily focused on demonstrating the substantial equivalence of the digital read system to the manual read, rather than measuring the improvement in human performance when assisted by the AI. The Virtuoso™ System is described as an "aid to the pathologist" and an "adjunctive computer-assisted methodology," but the study evaluates its standalone performance against manual reading, and its reproducibility, not its comparative effectiveness in improving reader accuracy or efficiency.

6. If a Standalone (Algorithm Only Without Human-in-the-Loop Performance) Was Done

No, a standalone (algorithm only) performance study was not done or reported. The device is described as an "aid to the pathologist" and the software explicitly "makes no independent interpretations of the data and requires competent human intervention for all steps in the process."
The Digital Read (DR) method involves the pathologist reviewing the digital images. The performance metrics (OPA, PPA, NPA) are based on the pathologist's interpretation using the digital system, not an automated algorithm's output directly.

7. The Type of Ground Truth Used

The primary ground truth used for the method comparison study was the expert's own manual microscopic assessment of the formalin-fixed, paraffin-embedded tissue slides, using a traditional microscope. This acts as the "reference manual method."
For the reproducibility and precision studies, the ground truth for comparison was the clinical score assigned by pathologists (either their own previous scores for intra-reader, or other pathologists' scores for inter-reader/inter-scanner).
The ground truth categories were defined as:
- Negative: PR score of 0 to 0.99% positive staining
- Positive: PR score of ≥1% positive staining
- For scanner precision: 0 – 0.99%, 1–10%, and ≥ 10% positive staining.
There is no mention of pathology or outcomes data being used as an independent, external ground truth beyond expert consensus.

8. The Sample Size for the Training Set

The document does not provide information on the sample size used for the training set for the Virtuoso™ System's software. The study focuses on the clinical validation of the device, implying that the algorithm development (training) phase was completed prior to these validation studies.

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

The document does not provide information on how the ground truth for the training set was established. This information is typically part of the device development process and is not always included in the 510(k) summary for validation studies.

Summary

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Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002

July 16, 2015

Ventana Medical Systems, Inc. Yassine Labiad, MS, RAC Regulatory Affairs Manager 203 Ravendale Drive Mountain View, CA 94043

Re: K142965

Trade/Device Name: Virtuoso™ System for IHC PR (1E2) using the VENTANA iScan HT Regulation Number: 21 CFR & 864.1860 Regulation Name: Immunohistochemistry reagents and kits Regulatory Class: Class II Product Code: OEO Dated: July 13, 2015 Received: July 14, 2015

Dear Mr. Labiad:

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. 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 801); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set

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forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

If you desire specific advice for your device on our labeling regulation (21 CFR Part 801), please contact the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/Resourcesfor You/Industry/default.htm. Also, please note the regulation entitled. "Misbranding by reference to premarket notification" (21CFR 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 the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.

You may obtain other general information on your responsibilities under the Act from the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.

Sincerely yours,

Reena Philip -S

Reena Philip, Ph.D. Director Division of Molecular Genetics and Pathology Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health

Enclosure

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Indications for Use

510(k) Number (if known) K142965

Device Name

Virtuoso™ System for IHC PR (1E2) using the VENTANA iScan HT

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)

PLEASE DO NOT WRITE BELOW THIS LINE - CONTINUE ON A SEPARATE PAGE IF NEEDED.

FOR FDA USE ONLY

Concurrence of Center for Devices and Radiological Health (CDRH) (Signature)

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SECTION 5 510(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 is K142965.

807.92 (a)(1):
Name:	Ventana Digital Pathology
Address:	203 Ravendale RdMountain View, CA 94043
Phone:	(408) 585-7949
FAX:	(408) 207-4299
Contact:	Mr. Yassine Labiad

807.92 (a)(2): Device name- trade name and common name, and classification

Trade name: Virtuoso™ System for IHC PR (1E2) using the VENTANA iScan HT
Common Name: IHC Digital pathology system for immunochemistry-stained slides
21 CFR § 864.1860- Immunohistochemistry reagents and kits Classifications:

807.92 (a)(3): Identification of the legally marketed predicate devices

This Virtuoso™ System for IHC PR (1E2) using the VENTANA iScan HT is substantially equivalent to its immediate predecessors cleared under K111869 on March 5, 2012, and K122143 on September 19, 2013. The only difference between the two is the use of a new scanner. The new iScan HT scanner is a powerful, high throughput bright field scanner that Ventana Medical Systems has decided to offer as an alternative to the iScan Coreo, which is a low to medium volume instrument currently employed for use with the Virtuoso™ System for IHC PR (1E2). Both the new and the predicate devices are digital pathology system for the consistent assessment of pathology interpretations using immunohistochemically stained slides (in this case. stained for PR expression), and both systems include slide scanner hardware, and software that both automates the procedural steps and performs the analyses.

807.92 (a)(4): Device Description

General Description

The Virtuoso™ System is an instrument-plus-software system designed to assist the qualified pathologist in the consistent assessment of protein expression in

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immunohistochemically stained histologic sections from formalin-fixed, paraffinembedded normal and neoplastic tissues. The system consists of a slide scanner, computer, monitor, keyboard, and mouse for specific immunohistochemical markers, and software with a Windows web browser-based user interface. Virtuoso is a web-based, end-to-end, digital pathology software solution that allows pathology laboratories to acquire, manage, view, analyze, share, and report digital images of pathology specimens. Using the Virtuoso software, the pathologist can view digital images, add annotations and generate reports.

The Virtuoso software is designed to complement the routine Software: workflow of a qualified pathologist in the review of immunohistochemically stained histologic slides. The software makes no independent interpretations of the data and requires competent human intervention for all steps in the process. Complete software information, following FDA's 2005 guidance document for software documentation required for regulatory submissions is being provided in Section 6 of this premarket notification.

Additional Materials Required:

Ventana CONFIRM™ anti-Progesterone Receptor (PR) (1E2) Rabbit . Monoclonal Primary Antibody
Reagents for visualization, such as DAB chromogen ●
Associated materials for completing immunohistochemical staining according ● to the appropriate package insert; this submission includes slides stained using both the Benchmark XT and Benchmark ULTRA stainers.
. Color printer if user wishes to print color copies

Device Quality Control

The quality of results depends on the laboratory following the quality control instructions recommended in the labeling of the immunohistochemistry (IHC) reagents.

Device Calibration:

The VENTANA iScan HT slide scanner is calibrated and verified at the manufacturing facility before shipment. Upon installation, calibrations are performed and verified by Roche Customer Support. No additional calibration or verification is required or performed by the end user. Annual preventive maintenance performed by Roche Customer Support is recommended to ensure that the VENTANA iScan HT slide scanner is operating as intended.

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Summary of Procedure

Samples are obtained as formalin-fixed, paraffin-embedded tissue blocks. Histologic sections are prepared and mounted onto glass slides. Slides are reacted with the PR (1E2) primary antibody and are then visualized using DAB. Prepared slides are loaded into the Virtuoso system scanner and scanned. The resulting digital images are reviewed by the pathologist on a computer monitor.

807.92 (a)(5): Intended Use

The Virtuoso™ system provides automated digital slide creation, management, analysis, and viewing. It is intended for in vitro diagnostic use as an aid to the pathologist in the display, detection, counting, review and classification of tissues and cells of clinical interest based on particular morphology, color, intensity, size, pattern and shape.

The Virtuoso™ System for IHC PR (1E2) using the VENTANA iScan HT is for the digital read application. This particular Virtuoso system is intended for use as an aid to the pathologist in the qualitative detection of progesterone receptor (PR) protein in formalin-fixed, paraffin-embedded normal and neoplastic tissue. This device is an accessory to Ventana Medical Systems, Inc. CONFIRM™ anti-Progesterone Receptor (PR) (1E2) Rabbit Monoclonal Primary Antibody assay. The CONFIRM™ anti-Progesterone Receptor (PR) (1E2) Rabbit Monoclonal Primary Antibody assay is indicated for use as an aid in the assessment of breast cancer patients for whom endocrine treatment is being considered (but is not the sole basis for treatment).

Note: The IHC PR (1E2) Digital Read application is an adjunctive computerassisted methodology for the qualified pathologist in the acquisition and interpretation of images from microscope glass slides of breast cancer specimens stained for the presence of PR protein. The accuracy of the test results depends on the quality of the immunohistochemical staining. It is the responsibility of a qualified pathologist to employ appropriate morphological studies and controls as specified in the instructions for the CONFIRM™ anti-Progesterone Receptor (PR) (1E2) Rabbit Monoclonal Primary Antibody used to assure the validity of the Virtuoso System for IHC PR Digital Read scores. The actual correlation of CONFIRMTM anti-PR antibody to clinical outcome has not been established. This device is intended for IHC slides stained on the BenchMark XT and BenchMark ULTRA stainers. For prescription use only.

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807.92 (a)(6): Technological Similarities and Differences to the Predicate Devices

The similarities and differences between the two test systems are described below.

	Predicate 1	Predicate 2	New Device
Characteristic	Virtuoso™ System for IHC PR(1E2)	Virtuoso™ System forIHC PR (1E2)	Virtuoso™ System for IHCPR (1E2)Using the VENTANA iScan HT
	[Benchmark ULTRA Stainer]K122143	[Benchmark XT Stainer]K111869
IntendedUse/Indicationsfor Use	The Virtuoso System providesautomated digital slide creation,management, analysis, andviewing. It is intended for IVDuse as an aid to the pathologist inthe display, detection, counting,review and classification oftissues and cells of clinicalinterest based on particularmorphology, color, size,intensity, pattern and shape.	SAME	The Virtuoso™ systemprovides automated digital slidecreation, management, analysis,and viewing. It is intended forin vitro diagnostic use as an aidto the pathologist in the display,detection, counting, review andclassification of tissues andcells of clinical interest basedon particular morphology,color, intensity, size, patternand shape.
	The Virtuoso™ System for IHCPR (1E2) is for digital read andimage analysis applications. Thisparticular Virtuoso system isintended for use as an aid to thepathologist in the detection andsemi-quantitative measurementof progesterone receptor (PR)protein in formalin-fixed,paraffin-embedded normal andneoplastic tissue. This device isan accessory to Ventana MedicalSystems, Inc. CONFIRMTM anti-Progesterone Receptor (PR)(1E2) Rabbit MonoclonalPrimary Antibody assay. TheCONFIRM™ anti-ProgesteroneReceptor (PR) (1E2) RabbitMonoclonal Primary Antibodyassay is indicated for use as anaid in the assessment of breast	SAME	and shape.The Virtuoso™ System for IHCPR (1E2) using the VENTANAiScan HT is for the digital readapplication. This particularVirtuoso system is intended foruse as an aid to the pathologistin the qualitative detection ofprogesterone receptor (PR)protein in formalin-fixed,paraffin-embedded normal andneoplastic tissue. This device isan accessory to VentanaMedical Systems, Inc.CONFIRM™ anti-ProgesteroneReceptor (PR) (1E2) RabbitMonoclonal Primary Antibodyassay. The CONFIRM™ anti-Progesterone Receptor (PR)(1E2) Rabbit MonoclonalPrimary Antibody assay is
	cancer patients for whomendocrine treatment is beingconsidered (but is not the solebasis for treatment).Note: The IHC PR (1E2) DigitalRead and Image Analysisapplications are adjunctivecomputer- assisted		indicated for use as an aid inthe assessment of breast cancerpatients for whom endocrinetreatment is being considered(but is not the sole basis fortreatment).Note: The IHC PR (1E2)
	Predicate 1	Predicate 2	New Device
Characteristic	Virtuoso™ System for IHC PR(1E2)[Benchmark ULTRA Stainer]K122143	Virtuoso™ System forIHC PR (1E2)[Benchmark XT Stainer]K111869	Virtuoso™ System for IHCPR (1E2)Using the VENTANA iScanHT
	methodologies for the qualifiedpathologist in the acquisition andmeasurement of images frommicroscope glass slides of breastcancer specimens stained for thepresence of PR protein. Thepathologist should verifyagreement with the ImageAnalysis software applicationscore. The accuracy of the testresults depends on the quality ofthe immunohistochemicalstaining. It is the responsibilityof a qualified pathologist toemploy appropriatemorphological studies andcontrols as specified in theinstructions for theCONFIRM™ anti-ProgesteroneReceptor (PR) (1E2) RabbitMonoclonal Primary Antibodyused to assure the validity of theVirtuoso System for IHC PRDigital Read and Image Analysisscores. The actual correlation ofCONFIRM™ anti-PR antibodyto clinical outcome has not beenestablished. This device isintended for IHC slides stainedon the BenchMark XT andBenchMark ULTRA stainers.For prescription use only.	Same	Digital Read application is anadjunctive computer-assistedmethodology for the qualifiedpathologist in the acquisitionand interpretation of imagesfrom microscope glass slides ofbreast cancer specimens stainedfor the presence of PR protein.The accuracy of the test resultsdepends on the quality of theimmunohistochemical staining.It is the responsibility of aqualified pathologist to employappropriate morphologicalstudies and controls asspecified in the instructions forthe CONFIRMTM anti-Progesterone Receptor (PR)(1E2) Rabbit MonoclonalPrimary Antibody used toassure the validity of theVirtuoso System for IHC PRDigital Read score. The actualcorrelation of CONFIRM™anti-PR antibody to clinicaloutcome has not beenestablished. This device isintended for IHC slides stainedon the BenchMark XT andBenchMark ULTRA stainers.For prescription use only.
Specimen Type	Formalin-fixed, paraffin-embedded tissue stained byimmunohistochemical technique	Same	Same
System Operation(Digital Read andImage Analysis)	Histologic observation by apathologist through the imageviewer and image analysissystems	Same	Histologic observation by apathologist through the imageviewer system.
Hardware andSoftware	Ventana iScan slide scanner,computer, color monitor,proprietary software for PR(1E2)	Same	Ventana iScan HT scanner,computer, color monitor,proprietary software for PR(1E2)
Platform	Mouse, keyboard, windows web	Same	Same
Characteristic	Predicate 1Virtuoso™ System for IHC PR(1E2)[Benchmark ULTRA Stainer]K122143	Predicate 2Virtuoso™ System forIHC PR (1E2)[Benchmark XT Stainer]K111869	New DeviceVirtuoso™ System for IHCPR (1E2)Using the VENTANA iScanHT
Components	browser.
Primary Antibody(Assay) Reagent	Ventana CONFIRM™ anti-Progesterone Receptor (PR)(1E2) Rabbit MonoclonalPrimary Antibody	Same	Same
AncillaryReagents/Stainers	DAB universal chromogen kits,Slides stained with BenchmarkULTRA stainer	DAB universal chromogenkits, slides stained withBenchmark XT stainer	DAB universal chromogen kits,slides stained with BenchmarkXT and ULTRA stainers
Localization ofIHC positive stain	Nucleus	Same	Same
Interpretation	Interpretation is performed bythe pathologist.	Same	Same

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A Member of the Roche Group

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A Member of the Roche Group

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807.92 (b)(1/2): Brief Description of Clinical Data (Non-clinical data N/A)

The Virtuosond System for IHC PR (1E2) using the VENTANA iScan HT was clinically validated via a method comparison (agreement/concordance) study and a precision and reproducibility study. The method comparison study evaluated overall system performance in terms of agreement between the reference manual method (with a traditional microscope) and the digital read (DR) application of the Virtuoso system. Two sets of statistical analyses were performed for the method comparison study: 1) Initial analysis: This analysis involved the primary endpoint analysis for each of the 3 pathologists participating in the study. The primary end points included the overall percent agreement (OPA), positive percent agreement (PPA), and negative percent agreement (NPA) between the Digital Read (DR) method and Manual Read (MR) method. 2) The additional analysis employed the Generalized Linear Mixed Model (GLMM) analysis methodology for the primary endpoint analysis. This additional analysis was associated with model-based results. which enabled appropriate estimation within a repeated-measures design of the average reader performance using the comparator device. The GLMM analysis included all 3 pathologists' data.

The reproducibility components of the studies involved the intrapathologist/inter-day reproducibility evaluation as well the inter-pathologist reproducibility of the DR Virtuoso application.

The Scanner precision study involved a subset of the clinical cases (n = 40, both stainers) that were scanned two more times, and also scanned three times with two different scanners at two separate locations. This study evaluated scanner precision for both inter-scanner precision and intra-scanner/inter-day precision.

The data from both method comparison and precision/reproducibility studies are summarized below.

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Method comparison (Agreement/Concordance)

Virtuoso Digital Read vs Manual Method

Each pathologist's Virtuoso digital read results were compared to their manual results. The data were categorized as "neg" and "pos" using the PR score of 0 to 0.99% positive staining to describe negative, and ≥1% positive staining to describe positive. The overall percent agreements using negative and positive, as defined above, across the three sites were: 98.7%, 88.9%, and 96.2%, respectively. The overall, negative, and positive percent agreements, with the 95% confidence intervals (CI) around the agreements are shown below.

Table: PR Clinical Status Agreement6550 Between DR and MR[6], By Site (Reader)

		Manual Read
		SITE 1(n =159)		SITE 2(n =162)		SITE 3(n =156)		Overall(n = 477)
		Pos	Neg	Pos	Neg	Pos	Neg	Pos	Neg
Digital Read	Pos	107	1	72	7	68	0	247	8
	Neg	1	50	11	72	6	82	18	204
	Total	108	51	83	79	74	82	265	212
Overall agreement (%)(95% CI) [d]		98.7(95.5-99.7)		88.9(83.1-92.9)		96.2(91.9-98.2)
Positive Agreement (%)(95% CI) [d]		99.1(94.9-99.8)		86.7(77.8-92.4)		91.9(83.4-96.2)
Negative agreement (%)(95% CI) [d]		98.0(89.7-99.7)		91.1(82.8-95.6)		100.0(95.5-100.0)

[a] Clinical status: positive = clinical scoring category of 1-10% or >10%; negative = clinical scoring category of 0-0.99%.

[b] Only cases with evaluable results are included this analysis. For a case result to be evaluable, the investigator must have assigned a valid clinical score (status) to the case and must have confirmed that the case morphology and background staining were acceptable and that the run controls were valid.

ld For sites that performed two independent MR scoring rounds, these data are from the first MR round.

[d] Two-sided 95% confidence interval calculated using the score method.

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Reproducibility

Intra-Pathologist/Inter-Day (pair-wise comparisons, Session 1 vs Session 2, a. Session 1 vs Session 3, Session 2 vs Session 3)

Digital Read

In the primary analysis of DR intra-reader reproducibility, the PR clinical status [negative (0-0.99%) or positive (≥1%)] was compared across three individual DR evaluations of the same set of 39 cases by the Reader. The 2 × 2 comparisons for the 3 reading sessions (reads) are shown in the following table with the respective agreement values. OPA rates for the 3 pairwise comparisons ranged from 89.7% to 97.4%. During the second and third reads for the DR reproducibility evaluation, Reader 3 investigator identified one case as unevaluable.

READ 1 vs READ 2			Read 2 Clinical Assessment			Read 1 vs Read 2 Agreement Rates
			Pos	Neg	Total	Rate	n/N	%	95% CI[b]
Read 1 ClinicalAssessment	Pos	19	1	20	APA	38/39	97.4	90.9-100.0
	Neg	0	19	19	ANA	38/39	97.4	90.7-100.0
	Total	19	20	39	OPA	38/39	97.4	86.8-99.5
READ 1 vs READ 3			Read 3 Clinical Assessment			Read 1 vs Read 3 Agreement Rates
			Pos	Neg	Total	Rate	n/N	%	95% CI[b]
Read 1 ClinicalAssessment	Pos	18	2	20	APA	36/40	90.0	78.0-97.9
	Neg	2	17	19	ANA	34/38	89.5	76.5-97.8
	Total	20	19	39	OPA	35/39	89.7	76.4-95.9
READ 2 vs READ 3			Read 3 Clinical Assessment			Read 2 vs Read 3 Agreement Rates
			Pos	Neg	Total	Rate	n/N	%	95% CI[b]
Read 2 ClinicalAssessment	Pos	18	1	19	APA	36/39	92.3	81.3-100.0
	Neg	2	18	20	ANA	36/39	92.3	81.3-100.0
	Total	20	19	39	OPA	36/39	92.3	79.7-97.3

PR clinical Status [3] Agreement Within Reader (Digital Read)

Clinical assessment (ie, PR status): positive = clinical score of 1—10%; negative = clinical score of 0 –

wo-sided 95% Cls for OPA of each read pair (not pooled for all read-pairs) are calculated using the score nethod. The other Cls are calculated using the percentile bootstrap method.

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b. Inter-Pathologist (pair-wise comparisons, Pathologist 1 vs Pathologist 2, Pathologist 1 vs Pathologist 3, Pathologist 2 vs Pathologist 3)
The agreement of PR clinical status between sites (readers) was examined separately for DR and MR. Each investigator's results for the PR clinical status [negative (0-0.99%) or positive (≥1%)] of the study cases were compared to those of another reader using the same reading method. The agreement rates and 95% CIs are shown in the following tables, respectively, for each reading method (DR and MR).

Manual Read

The three manual readings across the three pathologists (sites) were compared to each other. The reproducibility in the manual readings among the three pathologists (sites) is shown below, along with the 95% CIs. The OPA rates for the 3 pairwise comparisons ranged from 81.0% to 92.9%.

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SITE 1 vs. SITE 2					Site 1-vs-Site 2 Agreement Rates
	Site 2Clinical Assessment				Rate	n/N	%	95% CI [d] (%)
Site 1ClinicalAssessment	Pos	82	25	107	APA	164/190	86.3	80.7-91.2
	Neg	1	50	51	ANA	100/126	79.4	71.1-86.6
	Total	83	75	158	OPA	132/158	83.5	77.0-88.5
SITE 1 vs. SITE 3					Site 1-vs-Site 3 Agreement Rates
	Site 3Clinical Assessment				Rate	n/N	%	95% CI [d] (%)
Site 1ClinicalAssessment	Pos	74	29	103	APA	148/177	83.6	77.2-89.4
	Neg	0	50	50	ANA	100/129	77.5	68.8-85.1
	Total	74	79	153	OPA	124/153	81.0	74.1-86.5
SITE 2 vs. SITE 3					Site 2-vs-Site 3 Agreement Rates
	Site 3Clinical Assessment				Rate	n/N	%	95% CI [d] (%)
Site 2ClinicalAssessment	Pos	70	8	78	APA	140/151	92.7	88.0-96.7
	Neg	3	74	77	ANA	148/159	93.1	88.6-96.8
	Total	73	82	155	OPA	144/155	92.9	87.7-96.0

PR Clinical Status Agreement[8]] Between Sites (Readers): Manual Read

[a] Clinical status: positive = clinical scoring category of 1–10%; negative = clinical scoring category of 0 - 0.99%.

[c] Two-sided 95% confidence interval for OPA of each site (reader) pair (not pooled for all sites) are calculated using the score method. The other Cls are calculated using the percentile bootstrap method.

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Digital Read

The reproducibility in the Virtuoso digital readings among the three pathologists (sites) is shown below, along with the 95% Cls. The overall percent agreements ranged from 77.4% to 89.9%.

PR Clinical Status Agreement439) Between Sites (Readers): Digital Read

SITE 1 vs. SITE 2		Site 2Clinical Assessment			Site 1-vs-Site 2 Agreement Rates
		Pos	Neg	Total	Rate	n/N	%	95% CI [d] (%)
Site 1 ClinicalAssessment	Pos	79	29	108	APA	158/188	84.0	78.0-89.6
Site 1 ClinicalAssessment	Neg	1	50	51	ANA	100/130	76.9	68.8-84.6
Site 1 ClinicalAssessment	Total	80	79	159	OPA	129/159	81.1	74.3-86.5
SITE 1 vs. SITE 3		Site 3Clinical Assessment			Site 1-vs-Site 3 Agreement Rates
		Pos	Neg	Total	Rate	n/N	%	95% CI [d] (%)
Site 1 ClinicalAssessment	Pos	70	35	105	APA	140/175	80.0	73.4-86.3
Site 1 ClinicalAssessment	Neg	0	50	50	ANA	100/135	74.1	65.6-81.8
Site 1 ClinicalAssessment	Total	70	85	155	OPA	120/155	77.4	70.2-83.3
SITE 2 vs. SITE 3		Site 3Clinical Assessment			Site 2-vs-Site 3 Agreement Rates
		Pos	Neg	Total	Rate	n/N	%	95% CI [d] (%)
Site 2 ClinicalAssessment	Pos	65	11	76	APA	130/146	89.0	83.5-93.8
Site 2 ClinicalAssessment	Neg	5	77	82	ANA	154/170	90.6	85.7-94.7
Site 2 ClinicalAssessment	Total	70	88	158	OPA	142/158	89.9	84.2-93.7

[a] Clinical status: positive = clinical scoring category of 1–10% or ≥ 10%; negative = clinical scoring category of 0 – 0.99%.

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Scanner Precision

Forty (40) cases representing the useful categories of <1%, 1-10%, and 10% positive staining for PR were scanned on three different scanners at three different sites to assess inter-scanner precision, and three FOVs were captured (total = 120) and evaluated. Similarly, these same 40 cases were scanned on three different days by the same scanner, and the same FOVs were applied to the appropriate cases to assess intra-scanner/inter day precision.

Pairwise comparisons were performed between each of the three sites (interscanner), and between each of the three days (sessions, intra-scanner). The precision tables are found below.

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PR Clinical Scoring Category 19 Agreement Between Scanners (Between Sites): All FOVs Combined

SITE 1 vs SITE 2 [b]				Site 2 Clinical Scoring Category			OPA
			>10%	1–10%	0–0.99%	Total	n/N	%	95% CI[c]
Site 1 ClinicalScoringCategory	>10%	113	8	0	121	324/360	90.0	86.5-92.7
	1–10%	3	30	15	48
	0–0.99%	0	10	181	191
Total		116	48	196	360
SITE 1 vs SITE 3 [b]				Site 3 Clinical Scoring Category			OPA
			>10%	1–10%	0–0.99%	Total	n/N	%	95% CI[c]
Site 1 ClinicalScoringCategory	>10%	119	2	0	121	337/360	93.6	90.6-95.7
	1–10%	4	34	10	48
	0–0.99%	0	7	184	191
Total		123	43	194	360
SITE 2 vs SITE 3 [b]				Site 3 Clinical Scoring Category			OPA
			>10%	1–10%	0–0.99%	Total	n/N	%	95% CI[c]
Site 2 ClinicalScoringCategory	>10%	115	1	0	116	325/360	90.3	86.8-92.9
	1–10%	8	28	12	48
	0–0.99%	0	14	182	196
Total		123	43	194	360

[a] PR clinical scoring categories of: 0 – 0.99% (negative), 1–10% (positive), and ≥ 10% (positive). [b] This analysis is based on the results of pooling all days.

l Two-sided 95% Cls for OPA of each site (reader) pair (not pooled for all site-pairs) are calculated using the score method. Two-sided 95% Cls for OPA with all site-pairs pooled are calculated using the percentile bootstrap method.

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PR Clinical Scoring Category [3] Agreement Within Scanners (Between Days): All FOVs Combined

DAY 1 vs DAY 2 [b]				OPA
	>10%	1-10%	0-0.99%	Total	n/N	%	95% CI[c]
Day 1 ClinicalScoringCategory	>10%	117	4	0	121	332/360	92.2	89.0-94.6
	1-10%	1	33	11	45
	0-0.99%	1	11	182	194
	Total	119	48	193	360
DAY 1 vs DAY 3 [b]				OPA
	>10%	1-10%	0-0.99%	Total	n/N	%	95% CI[c]
Day 1 ClinicalScoringCategory	>10%	118	3	0	121	327/360	90.8	87.4-93.4
	1-10%	2	29	14	45
	0-0.99%	0	14	180	194
	Total	120	46	194	360
DAY 2 vs DAY 3 [b]				OPA
	>10%	1-10%	0-0.99%	Total	n/N	%	95% CI[c]
Day 2 ClinicalScoringCategory	>10%	115	3	1	119	327/360	90.8	87.4-93.4
	1-10%	5	31	12	48
	0-0.99%	0	12	181	193
	Total	120	46	194	360

[a] PR clinical scoring categories of: 0 – 0.99% (negative), 1–10% (positive), and ≥ 10% (positive). [b] This analysis is based on the results of pooling all FOVs and all sites.

[0 Two-sided 95% Cls for OPA of each day pair (not pooled for all day-pairs) are calculated using the score method. Two-sided 95% Cls for OPA with all day-pairs pooled are calculated using the percentile bootstrap method.

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807.92 (b)(3): Conclusions from Clinical Testing

Concordance, reproducibility, and precision studies were performed for the Virtuoso System for IHC PR (1E2) using the VENTANA iScan HT. The test system was shown to be as safe and effective (therefore substantially equivalent) as the predicate devices for its intended use.

Regulation Number and Section

§ 864.1860 Immunohistochemistry reagents and kits.

(a)
Identification. Immunohistochemistry test systems (IHC's) are in vitro diagnostic devices consisting of polyclonal or monoclonal antibodies labeled with directions for use and performance claims, which may be packaged with ancillary reagents in kits. Their intended use is to identify, by immunological techniques, antigens in tissues or cytologic specimens. Similar devices intended for use with flow cytometry devices are not considered IHC's.(b)
Classification of immunohistochemistry devices. (1) Class I (general controls). Except as described in paragraphs (b)(2) and (b)(3) of this section, these devices are exempt from the premarket notification requirements in part 807, subpart E of this chapter. This exemption applies to IHC's that provide the pathologist with adjunctive diagnostic information that may be incorporated into the pathologist's report, but that is not ordinarily reported to the clinician as an independent finding. These IHC's are used after the primary diagnosis of tumor (neoplasm) has been made by conventional histopathology using nonimmunologic histochemical stains, such as hematoxylin and eosin. Examples of class I IHC's are differentiation markers that are used as adjunctive tests to subclassify tumors, such as keratin.(2) Class II (special control, guidance document: “FDA Guidance for Submission of Immunohistochemistry Applications to the FDA,” Center for Devices and Radiologic Health, 1998). These IHC's are intended for the detection and/or measurement of certain target analytes in order to provide prognostic or predictive data that are not directly confirmed by routine histopathologic internal and external control specimens. These IHC's provide the pathologist with information that is ordinarily reported as independent diagnostic information to the ordering clinician, and the claims associated with these data are widely accepted and supported by valid scientific evidence. Examples of class II IHC's are those intended for semiquantitative measurement of an analyte, such as hormone receptors in breast cancer.
(3) Class III (premarket approval). IHC's intended for any use not described in paragraphs (b)(1) or (b)(2) of this section.
(c)
Date of PMA or notice of completion of a PDP is required. As of May 28, 1976, an approval under section 515 of the Federal Food, Drug, and Cosmetic Act is required for any device described in paragraph (b)(3) of this section before this device may be commercially distributed. See § 864.3.