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Development and use of a multiplex PCR system for the rapid screening of heat labile toxin I, heat stable toxin II and shiga-like toxin I and II genes of Escherichia coli in water

机译：Development and use of a multiplex pCR system for the rapid screening of heat labile toxin I, heat stable toxin II and shiga-like toxin I and II genes of Escherichia coli in water

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Enterotoxigenic Escherichia coli (ETEC) may produce heat-labile toxin (LT)I and LTII and heat-stable toxin (ST) I and STII, while shiga toxin producing E. coli (STEC) strains, including enterohaemorrhagic E. coli (EHEC), may produce shiga-like toxin (SLT) I and/or SLTII. Both ETEC and STEC are pathogenic to humans, pigs and cattle. As contamination of environmental water by any of these pathogenic E. coli cells is possible, a multiplex polymerase chain reaction (PCR) system for the rapid screening of LTI, STII, and SLTI and SLTII genes of E. coli was developed. The PCR primers used were the SLTI and SLTII genes specific primers developed by the present authors and the LTI and STII genes specific primers reported by other laboratories. The detection specificity of this multiplex PCR system was confirmed by PCR assay of ETEC, STEC and other E. coli cells as well as non-E. coli bacteria. Its detection limit was 10(2)-10(3) cfu each of the target cells per assay. When this multiplex PCR system was used for the rapid screening of LTI, STII ETEC and STEC in water samples such as tap, underground and lake waters, it was found that after the enrichment step, as few as 10(0) cells 100 ml(-1) of the water sample could be detected. Therefore, this PCR system could be used for the rapid monitoring of ETEC and/or STEC cells contaminating water samples.
机译：产肠毒素的大肠杆菌（ETEC）可能产生不耐热毒素（LT）I和LTII以及热稳定毒素（ST）I和STII，而产生志贺毒素的大肠杆菌（STEC）菌株，包括肠出血性大肠杆菌（EHEC）可能会产生志贺样毒素（SLT）I和/或SLTII。 ETEC和STEC均对人，猪和牛有致病性。由于这些致病性大肠杆菌细胞都可能污染环境水，因此开发了用于快速筛选大肠杆菌LTI，STII和SLTI和SLTII基因的多重聚合酶链反应（PCR）系统。所使用的PCR引物是本作者开发的SLTI和SLTII基因特异性引物，以及其他实验室报道的LTI和STII基因特异性引物。通过对ETEC，STEC和其他大肠杆菌细胞以及非E细胞进行PCR检测，证实了该多重PCR系统的检测特异性。大肠杆菌。每次检测每个靶细胞的检出限为10（2）-10（3）cfu。当将此多重PCR系统用于自来水，地下水和湖水等水样品中LTI，STII ETEC和STEC的快速筛选时，发现富集步骤后，100 ml（ -1）可以检测到水样。因此，该PCR系统可用于快速监测污染水样的ETEC和/或STEC细胞。

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    Tsen, H.Y.; Jian, L.Z.;

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  • 年度 2014
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