A general primer GP5+/GP6(+)-mediated PCR-enzyme immunoassay method for rapid detection of 14 high-risk and 6 low-risk human papillomavirus genotypes in cervical scrapings

J Clin Microbiol. 1997 Mar;35(3):791-5. doi: 10.1128/jcm.35.3.791-795.1997.

Abstract

Two cocktails of digoxigenin-labeled human papillomavirus (HPV) type-specific oligonucleotide probes and an enzyme immunoassay (EIA) were used as a basis to developed a group-specific detection method for 14 high-risk (types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68) and 6 low-risk (types 6, 11, 40, 42, 43, and 44) HPVs, following a general primer GP5+/bioGP6(+)-mediated PCR. The sensitivity of this high-risk/low-risk (HR/LR) HPV PCR-EIA ranged from 10 to 200 HPV copies, depending on the HPV type. Comparison of HR/LR HPV PCR-EIA with radioactive Southern blot hybridization using a general probe on the same PCR products derived from 417 cytomorphologically abnormal cervical scrapings resulted in an overall agreement of 96% between the two methods. Complete concordance between group-specific HR/LR detection and individual typing results for both single and multiple infections indicate the strong specificity of this HR/LR HPV PCR-EIA. Multiple infections could be predicted by comparing PCR-EIA optical density values of the cocktail probes with one of the individual oligonucleotide probes. This novel HR/LR PCR-EIA allows accurate and rapid identification of high-risk and low-risk HPV types in cervical scrapings and will facilitate HPV detection in HPV mass-screening programs.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Cervix Uteri / virology*
  • DNA Primers / genetics
  • Female
  • Genotype
  • Humans
  • Immunoenzyme Techniques* / statistics & numerical data
  • Mass Screening
  • Papillomaviridae / genetics*
  • Papillomaviridae / isolation & purification*
  • Papillomaviridae / pathogenicity
  • Papillomavirus Infections / complications
  • Papillomavirus Infections / virology
  • Polymerase Chain Reaction / methods*
  • Polymerase Chain Reaction / statistics & numerical data
  • Risk Factors
  • Sensitivity and Specificity
  • Tumor Virus Infections / complications
  • Tumor Virus Infections / virology
  • Uterine Cervical Diseases / complications
  • Uterine Cervical Diseases / virology
  • Uterine Cervical Neoplasms / etiology
  • Uterine Cervical Neoplasms / prevention & control
  • Uterine Cervical Neoplasms / virology
  • Virology / methods
  • Virology / statistics & numerical data

Substances

  • DNA Primers