Ultra view polymer detection kit

Manufactured by Roche

Sourced in Germany

The Roche Ultra View Polymer Detection Kit is a lab equipment product designed for the detection and visualization of target antigens in tissue samples. It provides a reliable and efficient method for immunohistochemical analysis.

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Lab products found in correlation

Ventana benchmark xt, by Roche (3 mentions) Benchmark xt autostainer, by Roche (2 mentions) Benchmark ultra autostainer, by Roche (1 mentions) Desmin, by Agilent Technologies (1 mentions) Myogenin, by Roche (1 mentions) Sox10, by Biocare Medical (1 mentions) Factor xiiia, by Cell Marque (1 mentions) Calponin, by Agilent Technologies (1 mentions) β catenin, by BD (1 mentions) Meca 79, by Santa Cruz Biotechnology (1 mentions) Ventana benchmark autostainer, by Roche (1 mentions) Rnascope hpv hr18 probe, by Advanced Cell Diagnostics (1 mentions)

8 protocols using ultra view polymer detection kit

Immunohistochemical Evaluation of p16 Expression

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Five-micron sections from the tissue blocks were evaluated by immunohistochemistry for expression of a biomarker of HPV E7 oncoprotein activity, the CDK-inhibitor p16. Sections of formalin-fixed and paraffin-embedded tissues were deparaffinized and subjected to antigen retrieval using 10 mM citrate buffer (92° C for 30 minutes). P16 expression was evaluated by use of a mouse monoclonal antibody against p16 (MTM Laboratories, Heidelberg, Germany) visualized using the Ultra view polymer detection kit (Ventana Medical Systems, Inc. Tucson, AZ) on a Ventana BenchmarkXT autostainer (Ventana). P16 expression was scored using a 4-tiered system: 0 = completely negative staining; 1 = focal staining (less than 20% of tumor cells); 2 = patchy staining (20 – 70% of tumor cells); 3 = diffuse staining (nuclear and cytoplasmic staining in greater than 70% of tumor cells). As a surrogate marker of HPV infection, only staining that was diffuse was regarded as positive for p16 overexpression.

Poling J., Ma X.J., Bui S., Luo Y., Li R., Koch W, & Westra W. (2014). Human papillomavirus (HPV) status of non-tobacco related squamous cell carcinomas of the lateral tongue. Oral oncology, 50(4), 306-310.

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Detecting MCPyV and HPV in OCC

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A tissue microarray was constructed using JHH OCC samples with available formalin-fixed, paraffin-embedded (FFPE) tissue blocks. Antigen retrieval was performed on deparaffinized, 4 micron sections using 10 mM citrate buffer at 92°C for 30 minutes. A mouse monoclonal antibody for MCPyV Large T-antigen (Santa Cruz Biotechnology, Dallas, TX; clone CM2B4; 1:500 dilution) was applied using a BenchMark Ultra autostainer (Ventana Medical Systems, Tucson, AZ). Signals were visualized using the UltraView polymer detection kit (Ventana). Staining was performed according to manufacturer’s instructions with a known positive case of Merkel cell carcinoma as a positive control and benign tonsillar tissue as a negative control. The presence of staining was assessed by a head and neck pathologist (LR).
Human papillomavirus (HPV) testing was performed on evaluable whole-slide specimens from all sites using p16 immunohistochemistry and HPV RNA in situ hybridization (ISH) for high-risk types as described elsewhere^{9 (link)}.

Windon M., Fakhry C., Rooper L., Ha P., Schoppy D., Miles B., Koch W., Vosler P., Eisele D, & D’Souza G. (2020). The role of age and Merkel cell polyomavirus in oral cavity cancers. Otolaryngology--head and neck surgery : official journal of American Academy of Otolaryngology-Head and Neck Surgery, 163(6), 1194-1197.

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Immunohistochemical Profiling of Rare Tumors

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Immunohistochemistry was performed on all cases, either at the time of original diagnosis or for the purposes this study. Whole-slide sections of formalin-fixed, paraffin embedded tumor tissue were cut at five-micron thickness, deparaffinized, and subjected to antigen retrieval using 10 mM citrate buffer at 92°C for 30 minutes. Immunohistochemistry was performed on all cases using antibodies for S100 (Ventana Medical Systems, Tucson, AZ), either Smooth Muscle Actin (Ventana) or Calponin (Dako, Carpinteria, CA), SOX10 (BioCare Medical, Concord, CA), Desmin (Dako), and β-catenin (BD Biosciences, San Jose, CA). In addition, for a subset of cases (n=6), PAX3 (Bioss Inc., Wolburn, MA) immunohistochemistry was also performed. Depending on tissue availability, immunohistochemistry was also performed in most cases using antibodies for myogenin (Ventana) and Factor XIIIa (Cell Marque, Rocklin, CA). All immunohistochemical signals were visualized using the Ultra view polymer detection kit (Ventana Medical Systems, Inc. Tucson, AZ) on a Ventana BenchmarkXT autostainer (Ventana). Staining was performed according to manufacturer's instructions in the presence of appropriate controls. “Focal” immunoreactivity was regarded as immunostaining in <10% of tumor cells.

Rooper L.M., Huang S.C., Antonescu C.R., Westra W.H, & Bishop J.A. (2016). Biphenotypic Sinonasal Sarcoma: an Expanded Immunoprofile Including Consistent Nuclear Beta-Catenin Positivity and Absence of SOX10 Expression. Human pathology, 55, 44-50.

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Tissue Microarray Immunohistochemistry Protocol

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Tissue microarrays (TMAs) were constructed for immunohistochemistry. The tissue cores (2 mm) from two representative areas were punched and placed into the recipient blocks using a manual TMA device (SuperBioChips Laboratories, Seoul, Korea). Immunohistochemistry was performed using a Ventana Benchmark Autostainer (Ventana Medical System, Tucson, AZ, USA). Briefly, the slides were de-paraffinized, and antigen retrieval was conducted using heat-induced (92 °C for 30 min) epitope retrieval with an MC1 solution (Ventana Medical System, Tucson, AZ, USA). Sections were incubated with primary antibodies: CD8 (1:100, C8/144B, Dako, Cambridge, UK), Foxp3 (1:100, 236A/E7, Abcam, Cambridge, UK), CD20 (1:600, L26, Dako, Cambridge, UK), and MECA-79 (1:200, Santa Cruz, Tucson, AZ, USA). The Ultraview Polymer Detection Kit (Ventana Medical System, Tucson, AZ, USA) was used for visualization, and the stained sections were counterstained with hematoxylin. Human tonsil tissue was used as the positive control tissue. A negative control was performed by replacing the primary antibody with normal serum.

Hong S.A., Hwang H.W., Kim M.K., Lee T.J., Yim K., Won H.S., Sun D.S., Kim E.Y, & Ko Y.H. (2020). High Endothelial Venule with Concomitant High CD8+ Tumor-Infiltrating Lymphocytes Is Associated with a Favorable Prognosis in Resected Gastric Cancer. Journal of Clinical Medicine, 9(8), 2628.

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Immune Cell Infiltrate Quantification

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5 μM tissue sections were stained with CD4 and CD8 using the Ultraview polymer detection kit (Ventana Medical Systems, Oro Valley, AZ) on a Ventana BenchmarkXT autostainer (Ventana Medical Systems, Oro Valley, AZ) under a standard protocol with isotype control antibodies by the Pathology Core Laboratory at the Johns Hopkins Hospital (Baltimore, MD). The intensity of the immune infiltrate was graded as low (≤5% TIL staining), or moderate to high (>5% staining). Of the 53 OTSCCs, the intensity of CD4+ staining could be assessed in 89% (47/53) of the samples and the intensity of CD8+ staining in 91% (48/53) of the samples. Of the PD-L1 negative samples, 91% (10/11) were scored for CD4+ and CD8+ staining. Of the PD-L1 positive samples, 88% (37/42) were scored for CD4+ staining and 90% (38/42) were characterized for CD8+ staining. 89% of samples had comparative information for CD4+ as compared to CD8+ (47/53) T cell frequencies. All data are found in Supplementary Table 1.

Mattox A.K., Lee J., Westra W.H., Pierce R.H., Ghossein R., Faquin W.C., Diefenbach T.J., Morris L.G., Lin D.T., Wirth L.J., Lefranc-Torres A., Ishida E., Chakravarty P.D., Johnson L., Zeng Y.C., Chen H., Poznansky M.C., Iyengar N.M, & Pai S.I. (2017). PD-1 expression in head and neck squamous cell carcinomas derives primarily from functionally anergic CD4+ TILs in the presence of PD-L1+ TAMs. Cancer research, 77(22), 6365-6374.

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Immunohistochemical Detection of HPV

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HPV status on the resected cancers was confirmed within the tumor by overexpression of p16 which is induced by HPV E7 onco-protein activity. Briefly, five micron sections of formalin fixed paraffin-embedded (FFPE) tissue were deparaffinized and subjected to antigen retrieval using 10 mM of citrate buffer (92 °C for 30 min). Expression of p16 was evaluated using a mouse monoclonal antibody against p16 (MTM Laboratories, Heidelberg, Germany), the visualization was performed using the Ultra View polymer detection kit (Ventana). p16 expression was scored as positive if strong and diffuse nuclear and cytoplasmic staining was present in >70% of the tumor (Fig. 1).

Smith D.F., Maleki Z., Coughlan D., Gooi Z., Akpeng B., Ogawa T., Bishop J.A., Frick K.D., Agrawal N., Gourin C.G., Ha P.K., Koch W.M., Richmon J.D., Westra W.H, & Pai S.I. (2014). Human papillomavirus status of head and neck cancer as determined in cytologic specimens using the hybrid-capture 2 assay. Oral oncology, 50(6), 600-604.

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P16 Immunohistochemistry for Tumor Analysis

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For p16 immunohistochemistry, 5-μm whole-slide tumor sections were deparaffinized and antigen retrieval was performed using 10 mM citrate butter at 92°C for 30 minutes. A mouse monoclonal antibody for p16 (clone E6H4; Ventana Medical Systems, Tucson, AZ; prediluted) was applied using a BenchMark XT autostainer (Ventana Medical Systems) with incubation for 8 minutes at 36°C, and signals were visualized using the ultraView polymer detection kit (Ventana Medical Systems). Staining was performed according to the manufacturer’s instructions in the presence of appropriate controls. p16 overexpression was defined as strong nuclear and cytoplasmic staining in > 70% of tumor cells.^{23 (link),24 (link)}

Rooper L.M., Windon M.J., Hernandez T., Miles B., Ha P.K., Ryan W.R., Van Zante A., Eisele D.W., D’Souza G., Fakhry C, & Westra W.H. (2020). HPV-positive Squamous Cell Carcinoma of the Larynx, Oral Cavity, and Hypopharynx: Clinicopathologic Characterization With Recognition of a Novel Warty Variant. The American journal of surgical pathology, 44(5), 691-702.

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HPV Tumor Status Evaluation in Oropharyngeal Cancers

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OPSCCs with available archival paraffin-embedded tumor specimens were tested for HPV tumor status. Hematoxylin and eosin sections from all cases were reviewed by a head and neck pathologist to confirm a diagnosis of squamous cell carcinoma. Whole-slide sections of formalin-fixed, paraffin-embedded primary tumors were cut at a 5-micron thickness and deparaffinized. Immunohistochemistry (IHC) was performed for p16 (clone INK4a; Ventana Medical Systems, Tucson, Arizona; prediluted) using the UltraView polymer detection kit (Ventana) on a BenchMark XT autostainer (Ventana). RNA in situ hybridization (ISH) for high-risk HPV E6/E7 messenger RNA (mRNA) was performed using the RNAscope HPV-HR18 Probe (Advanced Cell Diagnostics, Hayward, California), a single cocktail probe recognizing 18 high-risk HPV genotypes (16, 18, 26, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, 73, and 82), also on a BenchMark XT autostainer (Ventana). All staining was done according to manufacturers’ instructions in the presence of appropriate controls. p16 was considered positive in the presence of >70% diffuse nuclear and cytoplasmic positivity. HPV RNA ISH was scored as positive if multiple punctate brown signals could be identified in the cytoplasm or nucleus of >50% of tumor cells. OPSCCs were considered HPV related if they were both p16 positive and HPV RNA ISH positive.

Rettig E.M., Gooi Z., Bardin R., Bogale M., Rooper L., Acha E, & Koch W.M. (2019). Oral Human Papillomavirus Infection and Head and Neck Squamous Cell Carcinoma in Rural Northwest Cameroon. OTO Open, 3(1), 2473974X18818415.

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