DETECTION OF HUMAN PAPILLOMAVIRUS DNA BY IN-SITU HYBRIDIZATION AND PCR

In situ hybridization with non radioactive probes is attractive for hu man papillomaviruses (HPV) detection. Its sensitivity has been greatly improved by using different hybridization conditions, techniques for revealing the DNA-DNA hybrids and method of observation and various ce ll lines derived from human uterine carcinomas (CaSki, SiHa and HeLa c ells) which contain 500-600 copies of HPV DNA, 1-2 copies of HPV 16 an d 20-50 copies of HPV DNA 18, respectively. In situ gene amplification increased the detection of HPV DNA since hybridization spots were vis ible in SiHa cells on slides; a specific signal was observed in HeLa c ells in suspensions examined by flow cytometry. Confocal microscopy is an alternative method to in situ gene amplification since viral DNA i s detectable in SiHa cells with or without gene amplification. Thus, t he techniques used in this study are potentially useful for research o f single cellular genes.