Rapid detection of Escherichia coli virulence factor genes using multiplex real-time TaqMan® PCR assays

被引:31
作者
West, D. M.
Sprigings, K. A.
Cassar, C.
Wakeley, P. R.
Sawyer, J.
Davies, R. H.
机构
[1] Vet Labs Agcy, Technol Transfer Unit, Addlestone KT15 3NB, Surrey, England
[2] Vet Labs Agcy, Dept Food & Environm Safety, Addlestone KT15 3NB, Surrey, England
关键词
Escherichia coli; virulence factors; real-time PCR; multiplex PCR; diagnostics; validation;
D O I
10.1016/j.vetmic.2007.01.026
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Three multiplex real-time TaqMan (R) 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 H 7, 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 Mx3000p (TM) real-time PCR machine with the capacity to test up to 20 strains for 9 virulence genes in 1 h. (c) 2007 Elsevier B.V. All rights reserved.
引用
收藏
页码:323 / 331
页数:9
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