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Rapid Detection of Listeria monocytogenes in Raw Milk with Loop-Mediated Isothermal Amplification and Chemosensor

文献类型： 外文期刊

作者： Wang, Deguo 1 ; Zhang, Gaiping 1 ; Lu, Chengping 3 ; Deng, Ruiguang 1 ; Zhi, Aimin 1 ; Guo, Junqing 1 ; Zhao, Dong 1 ; Xu 1 ;

作者机构： 1.Henan Acad Agr Sci, Henan Key Lab Anim Immunol, Zhengzhou 450002, Peoples R China

2.Xuchang Univ, Chem & Chem Ind Coll, Xuchang 461000, Peoples R China

3.Nanjing Agr Univ, Anim Medial Coll, Nanjing 210095, Peoples R China

关键词： Listeria monocytogenes;loop-mediated isothermal amplification (LAMP);rhodamine-based dual chemosensor

期刊名称：JOURNAL OF FOOD SCIENCE （ 影响因子：3.167； 五年影响因子：3.376 ）

ISSN： 0022-1147

年卷期： 2011 年 76 卷 9 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 rhodamine-based dual chemosensor for much more efficient, field-friendly detection of Listeria monocytogenes. In this report, LAMP was performed at 63 degrees C for 10 min, followed by a rapid reaction of DNA amplification and the byproduct, pyrophosphate ion, with a rhodamine-based dual chemosensor and Cu2+ is visualized as a disappearance of red color. The detection limit of L. monocytogenes by the LAMP-chemosensor was 8 to 10 cells per reaction tube, 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 10560 reference method. The results showed that the LAMP-chemosensor method has the advantages of better sensitivity and speed and less dependence on equipment than the standard Polymerase Chain Reaction for specifically detecting low levels of L. monocytogenes DNA, and this can be useful in the field as a routine diagnostic tool.

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