Rapid detection of Escherichia coli virulence factor genes using multiplex real-time TaqMan PCR assays.
Three multiplex real-time TaqMan PCR assays were developed for the detection of Escherichia coli virulence factor genes in veterinary samples. Target virulence factors chosen were the fimbriae K88 (F4), K99 (F5), F41, F17, F18 and 987p (F6) and the toxins LT, STa and CDT IV. Detection of genes coding GAD were included in each assay as an internal control. These assays allow rapid identification of virulence factor genes using identical cycling conditions on an Mx3000Ptrade mark real-time PCR machine with the capacity to test up to 20 strains for 9 virulence genes in 1h.
West
D M
DM
Technology Transfer Unit, Veterinary Laboratories Agency, Woodham Lane, New Haw, Addlestone, Surrey, UK.
Sprigings
K A
KA
Cassar
C
C
Wakeley
P R
PR
Sawyer
J
J
Davies
R H
RH
D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis / Q000382:microbiology; D004269:DNA, Bacterial; D004926:Escherichia coli / Q000235:genetics / Q000472:pathogenicity*; D004927:Escherichia coli Infections / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D029968:Escherichia coli Proteins / Q000235:genetics*; D036781:Fimbriae Proteins / Q000235:genetics; D010861:Fimbriae, Bacterial / Q000235:genetics; D005784:Gene Amplification; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis / Q000382:microbiology; D037521:Virulence Factors / Q000235:genetics*
D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; D037521:Virulence Factors; D036781:Fimbriae Proteins