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HPV and p16 Testing in Oropharyngeal Squamous Cell Carcinoma

Methodology, Interpretation, and Significance

DNA In Situ Hybridization

Slide 23

March 2012

DNA in situ hybridization for HPV is often considered the gold standard for HPV detection. The biggest knock on HPV in situ hybridization is its low sensitivity; there are a sufficient number of false-negatives that should inspire you to question a negative result when the clinical and histologic parameters all seem to point towards an HPV-associated tumor. Several tumors with HPV DNA and E6/E7 mRNA have been negative by DNA in situ hybridization. The specificity of DNA in situ hybridization for HPV is remarkable, as it employs probes for select sequences (ie, low- or high-risk genotypes). Aside from artifactual staining, the presence of episomal or integrated staining patterns can be considered positive with confidence. Because this assay is interpreted using the light microscope, it not only allows for the detection of viral DNA, it provides insight into the localization of HPV DNA as well. That is, in situ hybridization permits the determination of episomal or fully integrated localization of HPV DNA. In situ hybridization is readily available in most laboratories and a fair number of laboratories belonging to institutions with smaller case volumes. Importantly, in situ hybridization gives no indication of biologic activity. That is, the presence of HPV DNA tells you little if anything, about its role in the initiation and/or maintenance of malignant transformation.

DNA In Situ Hybridization


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