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Development of Double Loop-Mediated Isothermal Amplification to Detect Listeria monocytogenes in Food

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Abstract

In this study, a double loop-mediated isothermal amplification (dLAMP) based on two target genes hlyA and iap was developed for the rapid detection of Listeria monocytogenes in food. The results revealed that the detection time and temperature of our dLAMP assay for L. monocytogenes were 15 min and 63 °C respectively, with a sensitivity of 10 fg DNA of L. monocytogenes per tube. While normal LAMP (nLAMP) of hlyA or iap was 100 fg DNA of L. monocytogenes per tube for 45 min and 63 °C. Furthermore, mineral oil and GoldViewII nucleic acid stain were chosen as the basic materials to develop a simple visualized identification of the positive samples. A total of 450 food samples were tested for L. monocytogenes using the dLAMP protocol developed in this study. The results showed that the accuracy of the dLAMP and the “gold standard” culture-biotechnical method were 100 % identical, suggesting that the modified dLAMP assay would provide a potential for detection of L. monocytogenes in food products.

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Acknowledgments

This research was supported by the Project of the National High Technology Research and Development Program of China (863 Program, No. 2012AA101601), Public Welfare Science and Technology Research Project of Hubei Province of P.R. China (2012DCA33001), and Science and Technology Research of Higher Education of Ningxia (NGY2012099).

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Authors and Affiliations

  1. Food Biotechnology and Food Safety Laboratory, College of Food Science and Technology, Huazhong Agricultural University, Wuhan, 430070, People’s Republic of China

    Rina Wu, Fusheng Chen & Xiaohong Wang

  2. College of Biological Science and Engineering, Beifang University of Nationalities, Yinchuan, 750021, People’s Republic of China

    Rina Wu

  3. Ningxia Center for Disease Control and Prevention, Yinchuan, 750021, People’s Republic of China

    Xiang Liu & Bangcheng Guo

Authors
  1. Rina Wu
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  2. Xiang Liu
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  3. Bangcheng Guo
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  4. Fusheng Chen
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  5. Xiaohong Wang
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Correspondence to Fusheng Chen.

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Wu, R., Liu, X., Guo, B. et al. Development of Double Loop-Mediated Isothermal Amplification to Detect Listeria monocytogenes in Food. Curr Microbiol 69, 839–845 (2014). https://doi.org/10.1007/s00284-014-0661-1

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  • DOI: https://doi.org/10.1007/s00284-014-0661-1

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