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Rapid and sensitive detection of Escherichia coli 0157 in raw milk by loop-mediated isothermal amplification combined with chemosensor

文献类型： 外文期刊

作者： Wang, Deguo 1 ; Chen, Junliang 2 ; Xiao, Fugang 1 ; Zhang, Gaiping 3 ; Huo, Guicheng 4 ;

作者机构： 1.Xuchang Univ, Food Sci & Engn Coll, Xuchang 461000, Peoples R China

2.Henan Univ Sicence & Coll, Food Sci & Engn Coll, Luoyang 471003, Peoples R China

3.Henan Acad Agr Sci, Henan Key Lab Anim Immunol, Zhengzhou 450002, Peoples R China

4.NE Agr Univ, Minist Educ China, Key Lab Dairy Sci, Harbin 150030, Peoples R China

关键词： Escherichia coli 0157 (detection with LAMP);LAMP (detection of Escherichia coli 0157)

期刊名称：MILCHWISSENSCHAFT-MILK SCIENCE INTERNATIONAL （ 影响因子：0.247； 五年影响因子：0.266 ）

ISSN： 0026-3788

年卷期： 2012 年 67 卷 4 期

页码：

收录情况： SCI

摘要： Loop-mediated isothermal amplification (LAMP) allows a rapid amplification of nucleic acids under isothermal conditions. It can be combined with a chemosensor for much more efficient, field-friendly detection of Escherichia coli 0157. In this report, LAMP was performed at 65 C for 10 min, followed by a rapid reaction of DNA amplification by product, pyrophosphate ion, with a chemosensor resulting in red disappearance. The detection limit of E. coli 0157 by the LAMP-Chemosensor was 3-5 copies, and the total assay time including 10 min for rapid DNA extraction was approximately 30 min. Data on naturally contaminated raw milk samples indicated that the LAMP method was highly specific and sensitive, giving 100% concordance with the ISO 16654:2001 reference method. The results showed that the LAMP-Chemosensor method had the advantages hierhin ich of better sensitivity and speed and less dependence on equipment than the standard (PCR) for specifically detecting low levels of E. coli 0157 DNA, and this can be useful in the field as a routine diagnostic tool.

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