Abstract
Background: Human papillomaviruses (HPVs) are classified into low-risk (LR) and high-risk (HR) types, depending on their association with the development of benign, potentially malignant or malignant lesions. Oral verrucal-papillary lesions (OVPL) are a distinct spectrum of epithelial lesions affecting the oral mucosa from benign to malignant. We aimed to determine the expression of HR-HPV in OVPLs and oral squamous cell carcinoma (OSCC) using real time quantitative polymerase chain reaction (qPCR) on formalin-fixed paraffin-embedded (FFPE) pathology samples.
Methods: FFPE tissue samples (n=43) were obtained and grouped into specific histopathological groups: squamous papilloma (n=10), verrucous hyperplasia (n=15), verrucous carcinoma (n=6), OSCC (n=10), and control cervical carcinoma (n=2). Detectable DNA was extracted using a GeneJET FFPE DNA Purification Kit. HR-HPV DNA was detected by a hybridisation primer/probe (Taqman) based qPCR Genesig ® kit.
Results: DNA was extracted from 40 of the 43 FFPE samples with a correlation of R 2 =0.8042 between DNA yield to the original tissue surface area. We recommend blocks should be of a confirmed diagnosis and contain >8 mm 2 of tissue. No HR-HPV DNA was detected in OVPL and OSCC groups using qPCR, while control DNA was detected.
Conclusions: We validated the GeneJET FFPE DNA Purification Kit and the GeneSig HR-HPV qPCR kit for FFPE tissue. In this pilot set of OVPL and OSCC samples from New Zealand no HR-HPV was detected.