Detection of Norwalk virus in the UK by the polymerase chain reaction.
We have developed a polymerase chain reaction for the detection of Norwalk virus using the published sequence of the virus RNA dependent RNA polymerase gene and have used this to clone and sequence this region of a virus from a UK outbreak. We have applied this method to a panel of UK Norwalk-like viruses using both Tet-z and Taq DNA polymerases and found that amplification produces a multiplicity of bands from stool samples. However, in combination with Southern blotting, Taq polymerase amplification detected virus in 13 of a panel of 30 clinical samples known to contain these viruses and also detected astroviruses in a mixed infection. Amplification using Tet-z DNA polymerase was less efficient (6/30) and detected predominantly viruses typed as UK type 2 by solid phase immune electron microscopy.
Willcocks
M M
MM
Division of Virology, New Medical School, Newcastle upon Tyne, UK.
Silcock
J G
JG
Carter
M J
MJ
D000595:Amino Acid Sequence; D001483:Base Sequence; D017250:Caliciviridae Infections / Q000453:epidemiology / Q000382:microbiology; D017931:DNA Primers; D004279:DNA, Viral / Q000235:genetics; D012321:DNA-Directed RNA Polymerases / Q000235:genetics; D004196:Disease Outbreaks; D005069:Evaluation Studies as Topic; D005759:Gastroenteritis / Q000453:epidemiology / Q000382:microbiology; D006801:Humans; D008969:Molecular Sequence Data; D009663:Norwalk virus / Q000201:enzymology / Q000235:genetics* / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D012680:Sensitivity and Specificity; D006113:United Kingdom / Q000453:epidemiology
D017931:DNA Primers; D004279:DNA, Viral; D012321:DNA-Directed RNA Polymerases