Failure to genotype herpes simplex virus by real-time PCR assay and melting curve analysis due to sequence variation within probe binding sites

J Clin Microbiol. 2003 May;41(5):2135-7. doi: 10.1128/JCM.41.5.2135-2137.2003.

Abstract

Real-time PCR with melting curve analysis of PCR products is a rapid procedure for detecting and genotyping herpes simplex virus (HSV). When testing mucocutaneous samples for HSV by a real-time PCR assay targeting the DNA polymerase gene, we found that some PCR products had atypical melting curves that did not conform to the expected melting temperatures for HSV type 1 or 2. Sequence analysis showed that these strains had base-pair mismatches over the probe binding sites. An alternative assay is required to type such atypical isolates.

MeSH terms

  • Base Pair Mismatch
  • Base Sequence
  • Binding Sites / genetics
  • DNA Probes / genetics
  • DNA-Directed DNA Polymerase / genetics
  • Genes, Viral
  • Genetic Variation
  • Genotype
  • Herpesvirus 1, Human / classification
  • Herpesvirus 1, Human / enzymology
  • Herpesvirus 1, Human / genetics
  • Herpesvirus 1, Human / isolation & purification
  • Herpesvirus 2, Human / classification
  • Herpesvirus 2, Human / enzymology
  • Herpesvirus 2, Human / genetics
  • Herpesvirus 2, Human / isolation & purification
  • Humans
  • Molecular Sequence Data
  • Nucleic Acid Denaturation
  • Polymerase Chain Reaction / methods
  • Sequence Homology, Nucleic Acid
  • Simplexvirus / classification
  • Simplexvirus / enzymology
  • Simplexvirus / genetics*
  • Simplexvirus / isolation & purification

Substances

  • DNA Probes
  • DNA-Directed DNA Polymerase