Pathology, research and practice · 2009
Sequence-based genotyping HPV L1 DNA and RNA transcripts in clinical specimens.
СтатьяПоддержка исследования: вне правительства СШАНазвание и аннотация: Перевод пока не подготовлен
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Satra Maria, Vamvakopoulou Dimitra N, Sioutopoulou Despina O, Kollia Panagoula, Kiritsaka Aspasia, Sotiriou Sotirios, Antonakopoulos Georgios, Alexandris Elias, Costantoulakis Pantelis, Vamvakopoulos Nicholas C
Аннотация
We developed a direct sequence-based genotyping method to detect single and multiple HPV L1 DNA and RNA types in genital and dermatological specimens. Our method couples PCR amplification of a highly conserved HPV L1 segment using a broad spectrum-generic primer cocktail mix with automated sequencing of amplified PCR products, followed by GenBank sorting of sequencing data. We genotyped 5 skin and 30 cervical HPV DNA-positive specimens using this method and established its first experimentally derived working cutoff value with the aid of commercial hybridization-based techniques. We suggest that sequence-based genotyping of appropriately amplified DNA and RNA products may serve as a primary HPV detection method in dermatological specimens. It can be applied as an all-purpose genotyping method for rare HPV types not detectable by commercial hybridization-based techniques and for sorting multiple HPV infections by order of prevalence.