Development of a LAMP assay for rapid detection of different intimin variants of attaching and effacing microbial pathogens
文献类型： 外文期刊
作者： Zhang Xue-han 1 ; Qing, Ye 1 ; Liu Ya-dong 2 ; Bin, Li 1 ; Renata, Ivanek 3 ; He Kong-wang 1 ;
作者机构： 1.Jiangsu Acad Agr Sci, Inst Vet Med, Natl Ctr Engn Res Vet Bioprod, Minist Agr,Key Lab Engn Res Vet Bioprod, Nanjing 210014, Jiangsu, Peoples R China
2.Nanjing Tech Bank Bioind Co Ltd, Nanjing 210000, Jiangsu, Peoples R China
3.Texas A&M Univ, Coll Vet Med & Biomed Sci, Dept Vet Integrat Biosci, College Stn, TX 77843 USA
期刊名称：JOURNAL OF MEDICAL MICROBIOLOGY （ 影响因子：2.472； 五年影响因子：2.836 ）
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收录情况： SCI
摘要： Intimin harboured by pathogenic Escherichia coli (E. coli) strains is a key virulence factor involved in host cell adherence and colonization. Twenty-seven intimin-encoding E. coli attaching and effacing (eae) 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 eae genes were designed using online software. The eae-LAMP assay was highly specific and detected all 27 tested eae variants; no cross-reactions were observed with genes from enterotoxigenic E. coli (ETEC), E. coli BL21, Salmonella, Shigella, Listeria monocytogenes, or Streptococcus suis type 2 (SS2). With the lowest detection limit of approximately 10 copies per reaction the eae-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 E. coli O157:H7 (eaeγ+), show that the eae-LAMP assay is a simple, rapid, sensitive and specific tool for detecting intimin variants from pathogenic strains of E. coli. The eae-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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