Evidence for transcriptionally active HPV oncogenes E6/E7 is regarded as the gold standard for presence of clinically relevant high-risk human papillomavirus (HPV), but detection of HPV E6 E7 mRNA can be challenging using conventional techniques. As a causal agent in head and neck squamous cell carcinoma (HNSCC), it is critical that the detection method enable pathologist review of tissue morphology and be of the highest specificity and sensitivity for accurate assessment of within the tissue microenvironment of FFPE specimens. RNAscope™ HPV Biomarker Detection Reagents and its proprietary “double Z” oligonucleotide probes specific for each subtype HPV E6/E7 mRNA enable high specificity detection of viral transcripts in routine FFPE tumor biopsies.

RNA ISH Assay Provides Highest Sensitivity and Specificity
RNAscope^® for in situ Detection of Transcriptionally Active Human Papillomavirus in Head and Neck Squamous Cell Carcinoma

Current methodologies for HPV testing include PCR-based amplification and DNA in situ Hybridization (ISH). PCR amplification of HPV DNA is more sensitive, but it is less specific than DNA ISH. Published studies (Bishop et. al., 2013, Upko et. al., 2011, and Schache et. al., 2013) indicate that RNAscope™-based ISH assay is more sensitive than DNA ISH in detecting HPV in Oropharyngeal Squamous Cell Carcinoma (OSCC) and results correlate well with p16 immunohistochemistry (IHC) staining (Table 1).  RNAscope™ ISH technology is an ideal platform giving the high sensitivity and specificity needed for detection of HPV biomarkers in Head and Neck Squamous Cell Carcinoma (HNSCC) samples.

TABLE 1. Using the current “gold-standard” qPCR as the reference method, RNAscope™ Technology demonstrated the best sensitivity and specificity for HPV status determination than existing methods (Schache et al, 2013).
Abbreviations: IHC=immunohistochemistry; ISH=in situ hybridization; NPV=negative predictive value; PPV=positive predictive value; qPCR=quantitative PCR