Characterization of "Norwalk-like viruses" and astroviruses by liquid hybridization assay.
"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.
Belliot
G M
GM
Division of Viral and Rickettsial Diseases, Viral Gastroenteritis Section (G-04), National Center for Infectious Diseases, Centers for Disease Control and Prevention, 1600 Clifton Road NE, Atlanta, GA 30333, USA.
Fankhauser
R L
RL
Monroe
S S
SS
D019350:Astroviridae Infections / Q000453:epidemiology / Q000821:virology; D017250:Caliciviridae Infections / Q000453:epidemiology / Q000821:virology; D004279:DNA, Viral / Q000378:metabolism; D004196:Disease Outbreaks; D005243:Feces / Q000821:virology; D005759:Gastroenteritis / Q000453:epidemiology / Q000821:virology; D005814:Genes, Viral; D005838:Genotype; D006801:Humans; D001257:Mamastrovirus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D009663:Norwalk virus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D009692:Nucleic Acid Heteroduplexes; D015345:Oligonucleotide Probes; D016366:Open Reading Frames / Q000235:genetics; D010802:Phylogeny; D012367:RNA, Viral / Q000032:analysis / Q000235:genetics / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013045:Species Specificity
D004279:DNA, Viral; D009692:Nucleic Acid Heteroduplexes; D015345:Oligonucleotide Probes; D012367:RNA, Viral