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

'Norvalk-like viruses' (NLVs) and human astroviruses are causative agents o f gastroenteritis in all age-groups. The typing of these agents is generall y done by nucleotide sequencing, blot hybridization, or enzyme immunoassay. These techniques are expensive. time-consuming, and sometimes require scar ce reagents, which limits the typing of NLVs and astroviruses to a few refe rence laboratories. This report describes a liquid hybridization assay that uses broadly reactive probes whose sequences are based on data from specim ens in collections available at CDC and GenBank. Two astrovirus genogroup-s pecific probes were designed and tested successfully on 26 wild strains fro m all serotypes. Fourteen GII and 16 GI representative NLV strains were typ ed without cross-hybridization by using P1B- and P2A-specific probes. descr ibed previously, and new P2B- and P1A-specific probes. Analysis of the spec ificity of the probes, the effect of the mismatches during hybridization, a nd the sensitivity of hybridization assay demonstrates this method to be a rapid and simple technique for molecular typing of NLVs and preliminary cha racterization of astroviruses. (C) 2001 Elsevier Science B.V. All rights re served.