1887

Abstract

Intimin harboured by pathogenic coli () strains is a key virulence factor involved in host cell adherence and colonization. Twenty-seven intimin-encoding attaching and effacing () gene variants have been reported according to their 3′ binding domain sequences. In our study, we developed a specific and sensitive loop-mediated isothermal amplification (LAMP) assay to detect all known intimin variants. Four primers specific for six regions of genes were designed using online software. The -LAMP assay was highly specific and detected all 27 tested variants; no cross-reactions were observed with genes from enterotoxigenic (ETEC), BL21, or type 2 (SS2). With the lowest detection limit of approximately 10 copies per reaction the -LAMP assay was 100 times more sensitive than conventional PCR. These results, and the results of tests involving food and faecal samples artificially contaminated with O157 : H7 (+), show that the -LAMP assay is a simple, rapid, sensitive and specific tool for detecting intimin variants from pathogenic strains of . The -LAMP assay has great potential for wider applications, not only in the laboratory but also in the field setting, as it does not require specialized equipment.

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2013-11-01
2019-10-14
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