A highly specific and sensitive loop-mediated isothermal amplification method for the detection of Escherichia coli O157:H7

被引：43

作者：

Ravan, Hadi ^{[1
]}

Amandadi, Mojdeh ^{[1
]}

Sanadgol, Nima ^{[2
,3
]}

机构：

[1] Shahid Bahonar Univ Kerman, Dept Biol, Fac Sci, Kerman, Iran

[2] Zabol Univ, Dept Biol, Fac Sci, Zabol, Iran

[3] Tehran Univ Med Sci, PSRC, Tehran, Iran

来源：

MICROBIAL PATHOGENESIS | 2016年 / 91卷

基金：

美国国家科学基金会;

关键词：

Loop-mediated isothermal amplification; LAMP assay; Escherichia coli O157:H7; Z3276; gene; POLYMERASE-CHAIN-REACTION; SEROTYPE O157-H7; RAPID DETECTION; MULTIPLEX PCR; GROUND-BEEF; IDENTIFICATION; SALMONELLA; ASSAY; GENE; STRAINS;

D O I：

10.1016/j.micpath.2015.12.011

中图分类号：

R392 [医学免疫学]; Q939.91 [免疫学];

学科分类号：

100102 ;

摘要：

E. coli O157:H7 is one of the most important foodborne pathogen that causes some human illnesses such as bloody diarrhea, hemolytic-uremic syndrome, and kidney failure. We developed a loop-mediated isothermal amplification (LAMP) assay with six special primers that target a highly specific 299-bp region of the 23276 gene for the detection of E. coli O157:H7. Among 117 bacterial strains tested in this study, positive results were only obtained from E. coli O157:H7 strains. The sensitivity level of the Z3276 LAMP assay was determined to be 5 CFU/reaction tube in pure bacterial culture. Moreover, the LAMP assay was successfully applied to artificially contaminated ground beef with a sensitivity level of 10(3) CFU/mL without pre-enrichment and 10 CFU/mL after a 4-h pre-enrichment. In conclusion, the present LAMP assay would be a useful and powerful tool for the rapid, sensitive, and specific diagnosis of E. coli O157:H7 strains in resource limited laboratories. (C) 2015 Elsevier Ltd. All rights reserved.

引用

页码：161 / 165

页数：5

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