Establishment of droplet digital PCR for quantitative detection of Listeria monocytogenes in food
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1.Science and Technology Research Center of China Customs, Beijing 100026, China;2.College of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029, China;3.China National Center for Food Safety Risk Assessment,Beijing 100021, China;4.Inner Mongolia Yili Industrial Group Co., Ltd, Beijing 100070, China

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R155

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    Abstract:

    Objective A droplet digital polymerase chain reaction (ddPCR) method was developed for the rapid quantitative detection of Listeria monocytogenes in food.Methods Specific primers and probes for Listeria monocytogenes were screened. The constant value effect of ddPCR method and plate counting method was compared through the detection of pure bacterial solution and artificially contaminated samples. The specificity, sensitivity and repeatability of ddPCR method were analyzed.Results The ddPCR had the characteristics of excellent specificity, sensitivity and repeatability in Listeria monocytogenes detection. The limit of detection (LOD) and the limit of quantification (LOQ) in pure bacterial solution were 136 CFU/mL. The LOQ in squid ring and sausage samples was 240 CFU/g and 155 CFU/g. The coefficient of variation of ddPCR was less than 25% at each gradient level, and the relative deviation of the logarithm of ddPCR and plate counting was less than 30%.Conclusion The established ddPCR method is rapid, accurate, sensitive and specific for the quantitative detection of Listeria monocytogenes in food.

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WEI Yongxin, MA Dan, DONG Yiyang, WEI Haiyan, LI Dan, ZHANG Ximeng, GUO Yunchang, LI Weiwei, PEI Xiaoyan, SONG Yueqian. Establishment of droplet digital PCR for quantitative detection of Listeria monocytogenes in food[J].中国食品卫生杂志,2022,34(5):937-942.

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History
  • Received:February 22,2022
  • Revised:
  • Adopted:
  • Online: December 01,2022
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