Detection and typing of human papillomavirus in cervical cancer in the Thai.

One hundred formalin-fixed paraffin embedded tissues with histopathologic diagnosed invasive cervical cancer (squamous cell carcinoma) were examined for the presence of HPV-DNA by polymerase chain reaction (PCR) using L1-consensus primers. The results indicated that 82 out of 100 (82%) samples were positive for HPV-DNA. Among the positive samples, 50 samples (61%) were typed by dot hybridization technique (DH). HPV-16 was the dominant type (42.68%), followed by HPV-18 (20.73%) and HPV-33 (3.66%). There were double infection of HPV-16 and 18 in 5 (6.1%) samples. None of HPV-6 and 11 were detected in this study. This finding suggests that HPV infection is an important etiologic factor for the development of cervical cancer especially the infection with high risk types, i.e., HPV-16 and 18.

Improvement of PCR detection of HPV-DNA using enhanced chemiluminescence system and dot hybridization.

Infection with human papillomavirus (HPV) is an important etiological factor in the development of cervical cancer; detection of the viral genome is of prognostic importance, particularly for preneoplastic lesions. Nowadays, polymerase chain reaction (PCR) technique is a method of choice for HPV detection because of its sensitivity. However, providing a reliable diagnostic test, both sensitivity and specificity of the test should be evaluated. In the present study, purified plasmid HPV-16 DNA and HeLa-DNA, containing HPV-18 DNA, were amplified by PCR using L1 consensus primers specific for HPV. The amplified product was then analysed by gel electrophoresis (GE) and dot hybridization (DH). The generic oligonucleotide probes (GPs) labelled with Enhanced Chemiluminescence 3' labelling kit (ECL) were used in DH. The sensitivity of PCR reaction after determining by GE was 1 copy per cell for purified plasmid HPV-16 DNA and 20-80 copies per cell for HeLa-DNA while determining by DH was only 0.1 and 2-8 copies per cell, respectively. Thus, the detection of amplified product by DH using enhanced chemiluminescence system improved not only the specificity but also the sensitivity of HPV detection at least 10-fold.