Intra-laboratory reproducibility of human papillomavirus identification in cervical specimens by a polymerase chain reaction-based assay

J Clin Virol. 2000 Dec;19(3):187-93. doi: 10.1016/s1386-6532(00)00142-6.

Abstract

Background: polymerase chain reaction (PCR)-based assays for human papillomavirus (HPV) sequences are in wide use in clinical and epidemiological studies. The reproducibility of these assays is not extensively studied.

Objectives: to estimate the intra-laboratory reproducibility of generic and type-specific HPV diagnoses by the MY09/MY11/HMB01 consensus L1 primer-based PCR assay.

Study design: systematically collected specimens (n=207) were masked and retested.

Results: when specimens negative in both initial and repeat assays were excluded from analysis, the diagnostic reproducibility was 98. 6% for beta-globin, 90.7% for generic HPV (any HPV type), and 76.9% for type-specific HPVs. The reproducibility of type-specific diagnosis increased with increase in signal strength in the hybridization reaction of the initial assay. When a specimen contained five or more HPV types in the initial assay, it was rare to identify all of the HPV types in the repeat assay.

Conclusions: the degree of reproducibility of the PCR diagnosis should be taken into account in the interpretation of HPV data in clinical and epidemiological studies.

Publication types

  • Comparative Study
  • Multicenter Study
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Cervix Uteri / virology*
  • DNA, Viral / analysis
  • Female
  • Genotype
  • Globins
  • Humans
  • Laboratories / standards*
  • Papillomaviridae / genetics
  • Papillomaviridae / isolation & purification*
  • Papillomavirus Infections / virology
  • Polymerase Chain Reaction / standards*
  • Reproducibility of Results
  • Tumor Virus Infections / virology

Substances

  • DNA, Viral
  • Globins