Typing of Human Papillomaviruses by Consensus Polymerase Chain Reaction and a Non-Radioactive Reverse Dot Blot Hybridization

J Virol Methods. 1994 Sep;49(2):129-39. doi: 10.1016/0166-0934(94)90037-x.

Abstract

A non-radioactive reverse dot blot hybridization method was developed for typing of human papillomavirus (HPV) consensus primer generated polymerase chain reaction (PCR) products in a single test. In the PCR biotin-14-dATP was incorporated into the amplified DNA, which was then used as a probe in hybridization with a membrane, on which different genital HPV types had been immobilized. Of cervical brush samples from women referred to a colposcopy clinic (n = 58) and from women attending a health control program (n = 14) which had been found positive by PCR with consensus HPV primers but negative using primers specific for the HPV types 6, 11, 16, 18, 31, 33 and 35, 25 (43%) and 3 (21%), respectively, could be typed by this method. The additional HPV types found were 34, 39, 40, 45, 52, 53, 56 and 58. Of the samples from the colposcopy clinic (n = 33) and the health control group (n = 11) which could not be typed, 23 and 5, respectively, showed HPV X which cross-hybridized with various HPV types under conditions of low stringency. It is possible to type by this fast and easy method consensus primer-generated PCR products of a wide range of HPV types or to verify the presence of HPV DNA of unknown types.

Publication types

  • Clinical Trial
  • Controlled Clinical Trial
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cervix Uteri / virology*
  • Cloning, Molecular / methods
  • Consensus Sequence
  • DNA / biosynthesis
  • DNA Primers
  • DNA, Viral / analysis
  • DNA, Viral / biosynthesis
  • Female
  • Humans
  • Nucleic Acid Hybridization
  • Papillomaviridae / classification*
  • Papillomaviridae / genetics*
  • Papillomaviridae / isolation & purification
  • Polymerase Chain Reaction / methods*
  • Reference Values

Substances

  • DNA Primers
  • DNA, Viral
  • DNA