Examination and evaluation of the usefulness of the in situ PCR detection technique for human papillomavirus in squamous cell carcinoma

Abstract

We developed a highly sensitive detection system for human papillomavirus(HPV) in order to elucidate the localization of HPV in formalin-fixed, paraffin-embedded sections from squamous cell carcinoma(SCC) using in situ PCR(consensus primers: MY 09 and MY 11). Various parameters such as thermal cycler apparatus, pepsin treatment, PCR reagent, stringency, PCR reaction condition, immunohistochemistry were examined. Under optimal conditions in situ PCR analysis revealed the specific presence of HPV DNA in all cases of SCC and their localizations were observed in the nuclei and cytoplasms of koilocytes, SCC cells and epithelial cells as forms of integrated HPV DNA and episomes. These results suggest that the establishment of the highly detection system for HPV brings to the development of carcinogenesis reserch much more.