Polymerase Chain Reaction for Human Picornaviruses

J Gen Virol. 1989 Dec;70 ( Pt 12):3261-8. doi: 10.1099/0022-1317-70-12-3261.

Abstract

We have used enzymic amplification of specific nucleic acid sequences followed by hybridization, for the rapid detection and typing of human picornaviruses after cell culture isolation. The test is based on the synthesis of cDNA, the polymerase chain reaction and the use of oligonucleotide probes. The primers were selected from the 5' non-coding region of the genome representing highly conserved regions. Sequences specific to enteroviruses and rhinoviruses were used as probes. The assay was able to identify all the picornavirus reference strains analysed and it was also possible to discriminate between enteroviruses and rhinoviruses by the hybridization procedure. When 29 picornavirus clinical isolates were analysed, all except one were detected by gel electrophoresis and a specific hybridization signal was obtained with all except three strains using the oligonucleotide probes.

MeSH terms

  • Base Sequence
  • DNA, Viral / analysis
  • Enterovirus / genetics
  • Gene Amplification*
  • Humans
  • Molecular Sequence Data
  • Nucleic Acid Hybridization
  • Oligonucleotide Probes
  • Picornaviridae / classification
  • Picornaviridae / genetics*
  • Picornaviridae / isolation & purification
  • Polymerase Chain Reaction*
  • Rhinovirus / genetics

Substances

  • DNA, Viral
  • Oligonucleotide Probes