The application of polymerase chain reaction to the detection of rotaviruses in faeces.
An assay protocol based on exploiting the polymerase chain reaction (PCR) for the detection of rotavirus in infected faeces is described. The assay is 100,000 times more sensitive than the standard electropherotype method that is widely used. It also gives a 5000-fold increase in sensitivity over the hybridisation based assay previously developed (Pedley and McCrae, 1984) and does not require the use of radioisotopes. The amplified product is a full length c-DNA copy of the gene encoding the major neutralisation antigen of the virus whose molecular cloning and sequence analysis will allow detailed information on the molecular basis of epidemiological variation to be rapidly collected.
Xu
L
L
School of Veterinary Science, University of Bristol, Langford, U.K.
Harbour
D
D
McCrae
M A
MA
D000818:Animals; D001483:Base Sequence; D002417:Cattle; D004279:DNA, Viral / Q000096:biosynthesis; D005243:Feces / Q000382:microbiology*; D005784:Gene Amplification; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012330:RNA, Double-Stranded / Q000032:analysis; D012401:Rotavirus / Q000302:isolation & purification*; D012400:Rotavirus Infections / Q000175:diagnosis*; D012680:Sensitivity and Specificity
D004279:DNA, Viral; D012330:RNA, Double-Stranded