Development of a Loop‐Mediated Isothermal Amplification Assay Based on lmo0460 Sequence for Detection of Listeria monocytogenes. Issue 3 (16th February 2015) - Liu, Zhanmin
Abstract Loop‐mediated isothermal amplification (LAMP) based on lmo0460 sequence in genome of Listeria monocytogenes (L. monocytogenes) was designed for detection of L. monocytogenes, which is an important foodborne kind of pathogenic bacteria causing human and animal disease. The primers set for lmo0460 sequence encoding membrane‐associated lipoprotein consist of four specific primers targeting six regions on specific fragment. The LAMP assay could be optimized and the optimum condition was in 50 min at 63°C. Amplification products were visualized by 2% agarose gel electrophoresis. Sensitivity of the LAMP assay for detection of L. monocytogenes in pure cultures was 1.7 cfu per reaction. The LAMP assay was 147‐fold higher sensitive than that of the conventional polymerase chain reaction (PCR) assay (251 cfu/reaction). Fifty chicken samples were detected for L. monocytogenes in the method, and the accuracy of LAMP was shown to be 100% when compared with culture biotechnical, while the PCR assay failed to detect L. monocytogenes in one of the positive samples. It is shown that LAMP assay based on lmo0460 sequence can be a highly sensitive, efficient and easy‐to‐perform useful detection tool for L. monocytogenes and will be a potential useful and powerful tool for detection foodborne pathogens. Practical Applications L . monocytogenes contamination in chicken samples has been reported. In developing countries, such as China, the LAMP assay is a suitable, useful tool for rapid detection of L. monocytogenes in food because it is more cost‐effective, simple and high sensitive than PCR.