Development of a Loop‐Mediated Isothermal Amplification Assay Based on lmo0460 Sequence for Detection of Listeria monocytogenes. Issue 3 (16th February 2015)

• Record Type: Journal Article
• Authors: Liu, Zhanmin; Zhu, Jiachao; Xia, Xueying; Wang, Lin; Yang, Cuiyun; Li, Xiaohong
• Is Part Of: Journal of food safety. Volume 35:Issue 3(2015:Jul.)
• Page Start: 362
• Page End: 369
• Publication Date: 2015-02-16
• Subjects: Food adulteration and inspection -- Periodicals; Food contamination -- Periodicals; Food -- Analysis -- Periodicals; Food -- Microbiology -- Periodicals; Pathogenic bacteria -- Periodicals; Food handling -- Periodicals; Food preservatives -- Periodicals; 664
• DOI: 10.1111/jfs.12183
• Languages: English
• ISSNs: 0149-6085

Abstract

<abstract abstract-type="main"> <title>Abstract</title> <sec id="jfs12183-sec-0001" sec-type="section"> <p>Loop‐mediated isothermal amplification (LAMP) based on lmo0460 sequence in genome of <italic>L</italic><italic>isteria monocytogenes</italic> (<italic>L</italic><italic>. monocytogenes</italic>) was designed for detection of <italic>L</italic><italic>. monocytogenes</italic>, 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 <italic>L</italic><italic>. monocytogenes</italic> 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 <italic>L</italic><italic>. monocytogenes</italic> 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 <italic>L</italic><italic>. monocytogenes</italic> 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 <italic>L</italic><italic>. monocytogenes</italic> and will be a potential useful and powerful tool for detection foodborne pathogens.</p> </sec> <sec id="jfs12183-sec-0002" sec-type="section"> <title>Practical Applications</title> <p> <italic>L</italic> <italic>. monocytogenes</italic> 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 <italic>L</italic><italic>. monocytogenes</italic> in food because it is more cost‐effective, simple and high sensitive than PCR.</p> </sec> </abstract>