VENTANA HER2 Dual ISH DNA Probe Cocktail assay

The VENTANA HER2 Dual ISH DNA Probe Cocktail is intended to determine HER2 gene status by enumeration of the ratio of the HER2 gene to Chromosome 17 by light microscopy. The HER2 and Chromosome 17 probes are detected using the VENTANA Silver ISH DNP Detection Kit and the VENTANA Red ISH DIG Detection Kit by a two-color chromogenic in situ hybridization (ISH) in formalin-fixed, paraffin-embedded human breast carcinoma tissue specimens, following staining on the BenchMark ULTRA instrument. The VENTANA HER2 Dual ISH DNA Probe Cocktail is indicated as an aid in the assessment of patients for whom Herceptin® (trastuzumab) is being considered. This product should be interpreted by a qualified pathologist in conjunction with histological examination, relevant clinical information, and proper controls. This product is intended for in vitro diagnostic (IVD) use.¹

The new VENTANA HER2 Dual ISH DNA Probe Cocktail has a new probe design using oligonucleotide (or oligo for short) probes, which is a more specific, shorter probe. In addition, it is to be used in conjunction with two new detection kits with improved sensitivity (VENTANA Silver ISH DNP Detection Kit and VENTANA Red ISH DIG Detection Kit). Additionally, all three of these new materials are in a new kit size. The VENTANA HER2 Dual ISH DNA Probe Cocktail is now a 30 test kit and both VENTANA Silver ISH DNP Detection Kit and VENTANA Red ISH DIG Detection Kit are now a 60 test kit.

The enhanced technology in the new assay shows improved performance in the following: New probe design allows for a shorter hybridization time which results in less failures and faster turnaround time. The hybridization step is shortened from approximately 6 hours to 1 hour, which decreases the chance for slide drying and associated speckling seen with previous generation HER2 SISH/Dual ISH assays. Also, the new detection kits, with hapten antibodies allow a more specific and sensitive reaction, resulting in brighter signal intensity making it easier and faster to read. Finally, the use of a part cell conditioning recovers more targets across a wider range of tissue pre-analytical conditions, and is largely responsible for the increase in first pass rates.

This product is an FDA Approved Class III PMA companion diagnostics product.

Yes, the INFORM HER2 DUAL ISH was discontinued January, 2022.

Only the ultraView Red ISH DIG Detection Kit was discontinued in January, 2022.

This assay is only validated with VENTANA Silver ISH DNP Detection Kit and VENTANA Red ISH DIG Detection Kit on BenchMark IHC/ISH instruments. Any deviation from the intended use stated in the package insert¹ is not recommended.

Pre-analytical conditions can influence the result of any ISH assay. Studies have found that the majority of “inconclusive” HER2 gene results by fluorescent in situ hybridization (FISH) relate to pre-analytical factors including “under” and “over” fixation⁵ and delay to fixation.⁶ Strict implementation of fixation procedures (e.g., a dedicated processor to ensure a minimum of 6 hours fixation) resulted in a 64% reduction in “inconclusive” cases from 10.8% failures to 3.4%⁵. To compensate for those variable pre analytical factors, this assay has been developed with certain selectable protocol steps for pre-treatment. Refer to the package insert and Interpretation Guide² for the recommendations about pre analytical conditions as well as other recommendations such as glass slide storage and handling, label placement and others.

Appropriately stained internal positive control nuclei are needed for interpreting results.² Roche recommends the use of HER2 Dual ISH 3-in-1 Xenograft Slides for initial assay installation and/or troubleshooting activities.

No, the VENTANA HER2 Dual ISH DNA Probe Cocktail assay runs on the BenchMark ULTRA instrument and can be read on a standard brightfield microscope. No additional platforms, fluorescence microscope or special dark room handling required.

Roche Tissue Diagnostics performed an internal study that showed no difference in staining when the VENTANA HE 600 system is used compared to the Sakura Tissue-Tek coverslipper.

Only fixation in 10% neutral buffered formalin (NBF) is recommended as some fixatives produce variable staining with ISH-based assays, including Bouin’s and Alcohol Formalin-Acetic Acid (AFA).

The principle of slide scoring has not changed. Roche created an Interpretation Guide² for this assay, with the intention of providing readers with a tool to facilitate HER2 gene status determination via interpretation of HER2 and Chr17 staining patterns using the VENTANA HER2 Dual ISH DNA Probe Cocktail. In the Interpretation Guide, you can find recommendations on how to determine if a slide is adequate for slide scoring and explanations about the scoring algorithm to determine HER2 gene status.

All our studies are performed in accordance with the FDA approved scoring and the assay was compared to the FDA approved FISH assay.

Using a dual ISH test allows for the visualization of several staining patterns, including Chromosome 17 polysomy.²

• No need for fluorescent microscope/oil immersion lens and darkroom
  • Fits in pathologists’ regular workflow at brightfield scope.
  • Her2 Dual ISH can be read alongside the H&E and other breast panel markers on the same case for easy comparison and correlation of findings.
  • No interference from tissue auto fluorescence or tissue marking dyes.
• Provides morphological context for the pathologist
  • Signals visualized at lower magnification allowing a pathologist to easily scan the entire tissue section to identify invasive carcinoma and tumor heterogeneity.
  • Counterstained with Hematoxylin II, like IHC slides, so morphologic features are preserved and visible.
  • Allows distinction between invasive and in situ carcinoma, identification of normal cells and easy comparison to H&E slide and IHC slides.
  • Normal cells provide a same slide internal control of the staining process so you can be confident in the results.
• Archivable results
  • Signals do not fade over time.
  • Allows other pathologists to consult and review difficult cases.
  • Slides can be stored long term alongside other H&E and IHC slides from the same case (no special storage needs).
  • Clear and intense signals, along with Hematoxylin II counterstain, are easily imaged with conventional slide scanners

Yes, included in the Package Insert is data showing that our test is highly concordant to FISH. The performance of these products is supported by significant data, including a clinical method comparison involving more than 600 breast cancer samples that analyzed the concordance of our assay with PathVysion FISH.¹