For in vitro diagnostic use. Others HER2 Dual ISH DNA Probe Cocktail Assay INFORM IVD Breast Carcinoma Gastric Carcinoma INFORM HER2 Dual ISH DNA Probe Cocktail Assay RTD000968 780-4422 INFORM HER2 Dual ISH DNA Probe Cocktail Assay 04015630970155 50 tests true INFORM HER2 DUAL ISH DNA PROBE CKTL US 05586640001 Not Available Reagents, kits en Ventana Medical Systems, Inc.’s (Ventana) INFORM 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. The HER2 and Chromosome 17 probes are detected by light microscopy using two color chromogenic in situ hybridization (ISH) in formalin-fixed, paraffin-embedded human breast cancer tissue specimens following staining on a BenchMark XT or BenchMark ULTRA instrument. The INFORM HER2 Dual ISH DNA Probe Cocktail is indicated as an aid in the assessment of breast cancer patients for whom Herceptin® (trastuzumab) or KADCYLA® (ado-trastuzumab emtansine) treatment is being considered.This product should be interpreted by a qualified reader in conjunction with histological examination, relevant clinical information, and proper controls.This reagent is intended for in vitro diagnostic (IVD) use. en The INFORM HER2 Dual ISH DNA Probe Cocktail (INFORM HER2 Dual ISH assay) is formulated for use with ultraView SISH DNP Detection Kit, ultraView Red ISH DIG Detection Kit, and accessory reagents on a BenchMark XT or ULTRA instrument.The detection kit contains a primary antibody and an enzyme-labeled secondary antibody conjugated to horseradish peroxidase (HRP) or alkaline phosphatase (AP) which is used as the chromogenic enzyme. During the Dual in situ hybridization (Dual ISH) staining process, DNP and DIG labeled probes are co-hybridized to their respective specific target DNA sequences within the cell nuclei. Detection of the DNP-labeled HER2 probe occurs first using the ultraView SISH DNP Detection Kit, which contains the following dispensers: a Rabbit anti-DNP Monoclonal Antibody, Multimer solution which contains a goat anti-rabbit secondary antibody conjugated to horseradish peroxidase (HRP), Chromogen A (Silver A), Chromogen B (Silver B) and Chromogen C (Silver C). Following incubation with primary Rabbit anti-DNP Antibody and then goat anti-rabbit HRP secondary antibody conjugate, the Silver In Situ Hybridization (SISH) reaction occurs. Briefly described, this reaction is driven by the sequential addition of Chromogens A (silver acetate), B (hydroquinone) and C (H2O2). Here, the silver ions (Ag+) are reduced by hydroquinone to metallic silver atoms (Ag0). This reaction is fueled by the substrate for HRP, hydrogen peroxide (Chromogen C). The silver precipitate is deposited in the nuclei and a single copy of the HER2 gene is visualized as a black dot.Figure 1 illustrates the SISH reaction.Following SISH detection for HER2, the DIG-labeled Chromosome 17 probe is detected with the ultraView Red ISH DIG Detection kit. This kit includes the following dispensers: a mouse anti-DIG monoclonal antibody, Red ISH Multimer solution which contains a goat anti-mouse IgG antibody conjugated to Alkaline Phosphatase (AP), pH Enhancer, Naphthol, and Fast Red. Following development of the SISH signal, the slide is incubated with the mouse anti-DIG antibody, which binds to the DIG hapten on the Chromosome 17 probe. The anti-hapten primary antibody is detected with the Multimer solution (goat anti-mouse IgG conjugated to AP enzyme). The slide is incubated with the pH Enhancer solution, which provides the proper salt components/concentrations and buffered pH for optimal AP enzyme performance. Next, naphthol phosphate is applied, which serves as the substrate for the AP enzyme (AP dephosphorylates naphthol). Fast Red, added to the slide next, combines with the dephosphorylated naphthol to form a red precipitate, which is readily visualized by light microscopy.Figure 2 illustrates the Red ISH reaction. The specimen is then counterstained with Hematoxylin II for interpretation by light microscopy.The staining protocol consists of numerous steps in which reagents are incubated for pre-determined times at specific temperatures. At the end of each incubation step, the BenchMark XT or ULTRA instrument washes the sections to remove unbound material and applies a liquid coverslip which minimizes the evaporation of the aqueous reagents from the slide. Results are interpreted using a light microscope and aid in the differential diagnosis of pathophysiological processes, which may or may not be associated with positive staining for the probe.Figure 1. SISH Reaction for HER2 Detection