使用多重实时TaqMan PCR检测法快速检测大肠杆菌毒力因子基因
Rapid detection of Escherichia coli virulence factor genes using multiplex real-time TaqMan PCR assays.
作者信息
West D M, Sprigings K A, Cassar C, Wakeley P R, Sawyer J, Davies R H
机构信息
Technology Transfer Unit, Veterinary Laboratories Agency, Woodham Lane, New Haw, Addlestone, Surrey, UK.
出版信息
Vet Microbiol. 2007 Jun 21;122(3-4):323-31. doi: 10.1016/j.vetmic.2007.01.026. Epub 2007 Feb 4.
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.
开发了三种多重实时TaqMan PCR检测方法，用于检测兽医样本中的大肠杆菌毒力因子基因。所选的目标毒力因子为菌毛K88（F4）、K99（F5）、F41、F17、F18和987p（F6）以及毒素LT、STa和CDT IV。每次检测都包含编码GAD的基因作为内部对照。这些检测方法能够在Mx3000P商标实时PCR仪上使用相同的循环条件快速鉴定毒力因子基因，1小时内最多可对20个菌株的9个毒力基因进行检测。
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