Multiplex PCR Assay for the Detection of Five Putative Virulence Genes Encoded in Verotoxigenic Escherichia coli Plasmids
Multiplex PCR Assay for the Detection of Five Putative Virulence Genes Encoded in Verotoxigenic Escherichia coli Plasmids
复制标题
DOI：
10.1007/s00284-011-9877-5
复制
发表时间：
2011-05-01
影响因子：
2.6
通讯作者：
Parma, A. E.
中科院分区：
文献类型：
--
作者：
Bustamante, A. V.;Sanso, A. M.;Parma, A. E.
目的是进行五价PCR检测VTEC质粒中编码的推定毒力基因，katP、espP、subA、stcE和ehxA。在检测中使用的五种特异性引物对不会相互干扰，并产生914、774、556、399和262 bp的扩增产物。随机选择属于20个血清型的39株菌株，以评估多种菌株中的多重性。本研究的结果表明，有可能进行同步扩增，并寻找公认的质粒编码的毒力标记，从不同的E。大肠杆菌血清型，并将该技术应用于遗传特性的E。从水库、食物或病人身上分离的大肠杆菌。这种互补技术是一种有用的工具，可用于检测流行病学研究的菌株间差异，并提供可能与人类感染风险相关的信息。
The aim was to perform a pentavalent PCR assay for the detection of putative virulence genes encoded in VTEC plasmids, katP, espP, subA, stcE, and ehxA. The five-specific primer pairs used in the assay do not interfere with each other and generate amplification products of 914, 774, 556, 399, and 262 bp. It was selected at random 39 strains belonged to 20 serotypes in order to evaluate the multiplex in a wide variety of strains. The results of this study indicate that it is possible to perform simultaneous amplification and search for recognized plasmid-encoded virulence markers from different E. coli serotypes and apply this technique to the genetic characterization of E. coli strains isolated from reservoirs, foods or patients. This complementary technique is a useful tool to detect interstrain differences for epidemiological studies and to provide information that could be related to the risk of human infection.