Characterization of "Norwalk-like viruses" and astroviruses by liquid hybridization assay

J Virol Methods. 2001 Feb;91(2):119-30. doi: 10.1016/s0166-0934(00)00254-8.

Abstract

"Norwalk-like viruses" (NLVs) and human astroviruses are causative agents of gastroenteritis in all age-groups. The typing of these agents is generally done by nucleotide sequencing, blot hybridization, or enzyme immunoassay. These techniques are expensive, time-consuming, and sometimes require scarce reagents, which limits the typing of NLVs and astroviruses to a few reference laboratories. This report describes a liquid hybridization assay that uses broadly reactive probes whose sequences are based on data from specimens in collections available at CDC and GenBank. Two astrovirus genogroup-specific probes were designed and tested successfully on 26 wild strains from all serotypes. Fourteen GII and 16 GI representative NLV strains were typed without cross-hybridization by using P1B- and P2A-specific probes, described previously, and new P2B- and P1A-specific probes. Analysis of the specificity of the probes, the effect of the mismatches during hybridization, and the sensitivity of hybridization assay demonstrates this method to be a rapid and simple technique for molecular typing of NLVs and preliminary characterization of astroviruses.

Publication types

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

MeSH terms

  • Astroviridae Infections / epidemiology
  • Astroviridae Infections / virology
  • Caliciviridae Infections / epidemiology
  • Caliciviridae Infections / virology
  • DNA, Viral / metabolism
  • Disease Outbreaks
  • Feces / virology
  • Gastroenteritis / epidemiology
  • Gastroenteritis / virology
  • Genes, Viral
  • Genotype
  • Humans
  • Mamastrovirus / classification*
  • Mamastrovirus / genetics
  • Mamastrovirus / isolation & purification
  • Norwalk virus / classification*
  • Norwalk virus / genetics
  • Norwalk virus / isolation & purification
  • Nucleic Acid Heteroduplexes
  • Oligonucleotide Probes
  • Open Reading Frames / genetics
  • Phylogeny
  • RNA, Viral / analysis
  • RNA, Viral / genetics
  • RNA, Viral / isolation & purification
  • Reverse Transcriptase Polymerase Chain Reaction*
  • Sensitivity and Specificity
  • Species Specificity

Substances

  • DNA, Viral
  • Nucleic Acid Heteroduplexes
  • Oligonucleotide Probes
  • RNA, Viral