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Pathol Res Pract ; 205(12): 863-9, 2009.
Article in English | MEDLINE | ID: mdl-19762162

ABSTRACT

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.


Subject(s)
Capsid Proteins/genetics , DNA, Viral/isolation & purification , Oncogene Proteins, Viral/genetics , Papillomavirus Infections/virology , RNA, Viral/isolation & purification , Sequence Analysis, DNA , Sequence Analysis, RNA , Uterine Cervical Dysplasia/virology , Uterine Cervical Neoplasms/virology , Warts/virology , Automation, Laboratory , Cervix Uteri/virology , Colposcopy , Databases, Genetic , Female , Genotype , Humans , Papillomavirus Infections/diagnosis , Polymerase Chain Reaction , Skin/virology , Uterine Cervical Neoplasms/diagnosis , Warts/diagnosis , Uterine Cervical Dysplasia/diagnosis
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