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[多重聚合酶链反应法快速检测食品中的肠出血性大肠杆菌]

[Rapid detection of enterohemorrhagic Escherichia coli in food with multiplex PCR assay].

作者信息

Wang J, Yang R, Guo Z, Qiu M

机构信息

China Import & Export Commodity Inspect Technology Institute, Beijing 100025, China.

出版信息

Wei Sheng Yan Jiu. 2001 Sep;30(5):310-2.

Abstract

Rapid detection of enterohemorrhagic Escherichia coli (EHEC) in food was achieved by using multiplex PCR assay. The assay was designed to amplify the 1109, 302 and 228 bp regions of corresponding virulence genes eaeA, hlyAB, slt1 and/or slt2, by using specific primers for each one reaction. In 60 EHEC and other strains, the above three virulence gene sequences were co-amplified from DNA isolated from 12 EHEC O157:H7 strains, 1 O26:H11 strain and 1 O111:H8 strain; eaeA sequence was amplified from DNA isolated from 2 EAEC strains; slt1/2 sequence was amplified from DNA isolated from 1 VTEC strain. No specific amplification product was produced from other 43 non-EHEC strains. For the detection of EHEC in food, results could be obtained within 8 hrs, and the sensitivity of assay was less than or equal to 1.6 cfu/g(ml) of food. The method was applied to 126 food samples, and EHEC was detected in 3 samples.

摘要

通过多重聚合酶链反应(PCR)检测法实现了食品中肠出血性大肠杆菌(EHEC)的快速检测。该检测法旨在通过使用针对每个反应的特异性引物,扩增相应毒力基因eaeA、hlyAB、slt1和/或slt2的1109、302和228bp区域。在60株EHEC及其他菌株中,从12株EHEC O157:H7菌株、1株O26:H11菌株和1株O111:H8菌株分离的DNA中共扩增出上述三种毒力基因序列;从2株肠集聚性大肠杆菌(EAEC)菌株分离的DNA中扩增出eaeA序列;从1株产志贺毒素大肠杆菌(VTEC)菌株分离的DNA中扩增出slt1/2序列。其他43株非EHEC菌株未产生特异性扩增产物。对于食品中EHEC的检测,可在8小时内获得结果,该检测法的灵敏度小于或等于每克(毫升)食品1.6个菌落形成单位(cfu)。该方法应用于126份食品样品,其中3份样品检测出EHEC。

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