食品中单核细胞增生李斯特菌微滴数字PCR定量检测方法建立
作者:
作者单位:

1.中国海关科学技术研究中心,北京 100026;2.北京化工大学生命科学与技术学院,北京 100029;3.国家食品安全风险评估中心,北京 100021;4.内蒙古伊利实业集团股份有限公司,北京 100070

作者简介:

魏咏新 女 工程师 研究方向为食品安全微生物检测 E-mail:657799307@qq.com

通讯作者:

宋悦谦 男 工程师 研究方向为预防医学 E-mail:13436500493@139.com

中图分类号:

R155

基金项目:

海关总署科研项目(2021HK215);国家质量监督检验检疫总局科研项目(2017IK176)


Establishment of droplet digital PCR for quantitative detection of Listeria monocytogenes in food
Author:
Affiliation:

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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    摘要:

    目的 建立食品中单核细胞增生李斯特氏菌的微滴式数字聚合酶链式反应(ddPCR)快速定量检测方法。方法 筛选单核细胞增生李斯特氏菌的特异性引物和探针,通过对目标菌纯菌液及人工污染样品的检测,比较ddPCR方法和平板计数法的定值效果,对ddPCR结果进行特异性、灵敏性和重复性分析。结果 本研究建立的单核细胞增生李斯特氏菌ddPCR检测方法具有良好的特异性、灵敏性和重复性。单核细胞增生李斯特氏菌纯菌液中定量限(LOQ)和检出限(LOD)均为136 CFU/mL,在鱿鱼圈和香肠样品中定量限分别为240 CFU/g和155 CFU/g。ddPCR在各梯度水平上变异系数均小于25%,ddPCR和平板计数定值对数值相对偏差均小于30%。结论 本研究建立的ddPCR方法能够快速、准确、灵敏、特异地定量检测食品中单核细胞增生李斯特氏菌。

    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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魏咏新,马丹,董益阳,魏海燕,李丹,张西萌,郭云昌,李薇薇,裴晓燕,宋悦谦.食品中单核细胞增生李斯特菌微滴数字PCR定量检测方法建立[J].中国食品卫生杂志,2022,34(5):937-942.

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  • 收稿日期:2022-02-22
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  • 在线发布日期: 2022-12-01
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