Vendruscolo J W, Waldrich T L, Saikawa G I A, Pelayo J S, Kobayashi R K T, Nakazato G, Rocha S P D
Departamento de Microbiologia, Centro de Ciências Biológicas, , , Brasil.
Departamento de Microbiologia, Centro de Ciências Biológicas, , , Brasil
Genet Mol Res. 2017 Aug 17;16(3):gmr-16-03-gmr.16039760. doi: 10.4238/gmr16039760.
Diarrhea is considered the second most common cause of infant mortality worldwide. The disease can be caused by many different pathogens, including diarrheagenic Escherichia coli (DEC), which includes the pathotypes enterotoxigenic E. coli (ETEC), enteroinvasive E. coli (EIEC), enteroaggregative E. coli (EAEC), Shiga toxin-producing E. coli (STEC), and enteropathogenic E. coli (EPEC). To develop a multiplex PCR system for the safe and accurate identification of the five main pathotypes of DEC, seven pairs of primers were determined for the following genes: aaiC, escV, bfpA, ipaH, elt, stx1, and stx2. To validate the system, 413 isolates from different sources (water and both animal and human stool) were analyzed that had been characterized previously. The sensitivity data were grouped by pathotype, in which 92.7% of the atypical EPEC were correlated, as were 92.8% of the STEC, 91.35% of the EAEC, and 100% of the typical EPEC, ETEC, and EIEC. These findings indicate that it is possible to detect the major five pathotypes of DEC from different sources, which can aid in determining the epidemiology of diarrhea with a low cost, high sensitivity and specificity, and the easy and safe viewing of the resulting PCR products.
腹泻被认为是全球婴儿死亡的第二大常见原因。该疾病可由许多不同的病原体引起，包括致泻性大肠杆菌（DEC），其中包括产肠毒素大肠杆菌（ETEC）、侵袭性大肠杆菌（EIEC）、聚集性大肠杆菌（EAEC）、产志贺毒素大肠杆菌（STEC）和致病性大肠杆菌（EPEC）等致病型。为开发一种用于安全准确鉴定DEC五种主要致病型的多重PCR系统，针对以下基因确定了七对引物：aaiC、escV、bfpA、ipaH、elt、stx1和stx2。为验证该系统，对先前已鉴定的来自不同来源（水以及动物和人类粪便）的413株分离株进行了分析。敏感性数据按致病型分组，其中92.7%的非典型EPEC、92.8%的STEC、91.35%的EAEC以及100%的典型EPEC、ETEC和EIEC具有相关性。这些结果表明，有可能从不同来源检测DEC的五种主要致病型，这有助于以低成本、高灵敏度和特异性确定腹泻的流行病学情况，并且能够轻松安全地观察PCR产物结果。
