[Broad-spectrum PCR assay for detection of human papillomaviruses in oropharyngeal cancer].
OBJECTIVE
The multitude of studies yielding varying results concerning analysis of the prevalence of human papillomavirus in squamous cell carcinoma of the head and neck gives emphasis to the different methodologies applied. Typical histological changes i. e. koilocytosis, hyperkeratosis and parakeratosis indicating HPV infection are frequently observed, but detection of the virus often fails.


METHODS
61 specimens of oropharyngeal squamous carcinomas were investigated at the time of primary diagnosis as well as after radiation or combined radio-chemotherapy. Aim of the present study was to establish a molecular detection system including a broad spectrum of different HPV-types. This could be achieved by applying so-called degenerated consensus primers. By using voluntarily altered base-pair sequences in a highly conserved region of the HPV genome, the oligonucleotide primers were sensitive to the huge sequence variance of different HPV-types. Furthermore, sensitivity of the primary PCR was enhanced by applying a subsequent nested PCR.


RESULTS
In 16 biopsies (26,2 %) investigated (n = 61) a positive signal was noted after at least one PCR assay. In seven specimens direct sequencing was successful.


CONCLUSIONS
The present study demonstrates that the established broad-spectrum PCR is a reliable method for detection of HPV-DNA.
von Winterfeld
F
F
Universitäts-Hals-Nasen-Ohren-Klinik, Universität Duisburg-Essen.
Fischer
M
M
D001706:Biopsy; D015344:DNA Probes, HPV / Q000235:genetics*; D006801:Humans; D009959:Oropharyngeal Neoplasms / Q000473:pathology / Q000821:virology*; D009960:Oropharynx / Q000473:pathology / Q000821:virology; D030361:Papillomavirus Infections / Q000473:pathology / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011237:Predictive Value of Tests; D017422:Sequence Analysis, DNA
D015344:DNA Probes, HPV