生食动物性水产品中单增李斯特氏菌TaqMan实时荧光PCR检测法和国标法的比较研究

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Comparative Study Between TaqMan Real-Time Fluorescence PCR Assay and National Standard Method for the Detection of Listeria monocytogenes in Raw Animal Aquatic Products

LIUAifang，ZHENG Ermei，WANGHanzheng，MAO Zhanhua， ZHU Weizhi （Zhejiang Gongzheng Testing Center， Hangzhou 31oooo， China）

Abstract： Objective： To systematically compare the technical characteristics of TaqMan real-time fluorescent PCR and national standard assays for the detection of Listeria monocytogenes in raw animal aquatic products，in order to optimize the detection processand improve the effciency of foodbome pathogen monitoring.Method：The optimal primer probe combinations of the TaqMan detection system were screened by specificity experiments to validate its sensitivityand detection stability; five types ofcommercially available raw aquatic products （mud snail， ready-to-eat jellyfish threads，choking crab，Arctic shrimp，and salmon） were detected toevaluate the diferences in detection eficacy between the two methods.Result： The optimal primer probe combination for LIS2 was obtained by screening，and its detection sensitivity reached 6×102 CFU mL-1 ，and the 20-day stability test showed that the coefficient of variation of the Ct value of the 6×103 CFU mL-1 bacterial suspension was 1.89% ，which demonstrated good stability.In20 commercially available samples，the positive detection rates were identical for both methods， and the TaqMan method took up to 50h less time than the national standard method for a single sample，but relied on nucleic acid amplification equipment. Conclusion： TaqMan real-time fluorescent PCR is suitable for the primary screening of large quantities of samples，and it is recommended to be combined with phage lysis technology to achievelive bacteria detection.The national standard method was used as a confirmation method to review and evaluate the positive samples to ensure that the detection was inaccordance with the regulations.The studyconfirmed that the establishment of a joint detection system of“molecular preliminary screening-culture confirmation”can improve the detection efficiency and provide data support for the standardization of food microbiological rapid detection technology and the safety supervision of raw aquatic products.

Keywords： Listeria monocytogenes; TaqMan real-time fluorescent PCR; raw animal aquatic products; national standard method

在全球饮食文化相互渗透、不断交融的时代浪潮中，刺身、牡蛎、生腌海鲜等生食动物性水产品，以其别具一格的风味和口感，在消费市场中迅速走红，深受广大消费者喜爱。（剩余8876字）