Sequence-based genotyping HPV L1 DNA and RNA transcripts in clinical specimens

Pathol Res Pract. 2009;205(12):863-9. doi: 10.1016/j.prp.2009.07.020. Epub 2009 Sep 16.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

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

Substances

  • Capsid Proteins
  • DNA, Viral
  • HPV L1 protein, Human papillomavirus
  • Oncogene Proteins, Viral
  • RNA, Viral