Detection of high risk human papillomavirus in routine cervical smears: strategy for screening.

Abstract

Aim: To develop a methodology for direct detection of high risk human papillomavirus (HPV) Recently the association between HPV and cervical neoplasia has been questioned because HPV sequences are demonstrable using the polymerase chain reaction (PCR) in cervical samples from a high proportion of normal women.`2 However, interpretation of the data has been problematic due to inconsistencies in methodology and results from different laboratories. Estimates of the prevalence of HPV infection in both normal and abnormal cervical epithelial cells have varied widely."2 The possibility of contamination with exogenous HPV sequences is difficult to exclude when methods are used which require destruction of cellular morphology. In situ hybridisation has the advantage that cellular localisation of sequences can be performed microscopically and it excludes the possibility of contamination. Isotopic detection of HPV in cervical smears has been previously described'3 and non-isotopic in situ hybridisation (NISH) has been performed on cell samples collected by cervicovaginal lavage.'4 These methods are not, however, applicable to cervical smears collected in the routine way. Methods PROBESGenomic probes for HPV 6, i 115, 16

Human Papilloma Virus 16 E6 RNA Interference Enhances Cisplatin and Death Receptor-Mediated Apoptosis in Human Cervical Carcinoma Cells

“…1). We tested several HPV16 E6 antibodies as described recently (Sima et al, 2008) but were unable to detect HPV16 E6 in SiHa cells, which are known to have extremely low copies of HPV16 (Herrington et al, 1992). Therefore, p53 protein levels were used as a functional readout for the efficacy of HPV16 E6 down-regulation (Sima et al, 2008;Yamato et al, 2008;Liu et al, 2009).…”

Detection of integrated high risk human papillomavirus in adenoid cystic carcinoma of the uterine cervix.

“…Methods-Eleven archival, paraffin wax embedded specimens were analysed by non-isotopic in situ hybridisation (NISH) for HPV types 6,11,16,18,31, and 33 using digoxigenin labelled probes. The polymerase chain reaction (PCR) was carried out on each of the cases using consensus primers to HPV.…”

“…'0 Certain types, such as HPV 6 and 1 1, are regarded as low risk as these types are associated predominantly with genital condylomata and low grade cervical dysplasia. Other types, such as 16 and 18 and to a lesser extent 31,33,35,45,51,52, and 56, however, are regarded as high risk viruses because of their well recognised association with high grade cervical intraepithelial neoplasia and invasive carcinoma." Integration of high risk HPV DNA into the human genome is a crucial step in the evolution of HPV induced cervical cancers.…”

Detection of human papillomavirus in matched cervical smears and biopsy specimens by non-isotopic in situ hybridisation.

“…Methods: HPV samples were assayed individually by NISH with digoxigenin labelled probes (HPV6, 11,16,18, and 33) on routinely collected paraffin wax embedded cervical biopsy specimens and for high risk HPVs with a cocktail of similarly labelled probes (HPV16, 18, 33) on matched smears. These were taken at the same colposcopic examination from 32 patients investigated for an abnormal cervical Papinacolau (PAP) stained smear.…”