多重PCR-膜芯片技术快速检测饲料中6种致病微生物  

Simultaneous detection of six pathogenic microorganisms in feed by PCR-membrane chip assay

作　　者：魏书林[1] 李文静 宋荣[1] 李丽蓓[1] 王培龙[1] 冯超林 张维[1] WEI Shulin;LI Wenjing;SONG Rong;LI Libei;WANG Peilong;FENG Chaolin;ZHANG Wei(Institute of Quality Standard and Testing Technology for Agro-Products,CAAS,Beijing 100081,China;Hubei Provincial Veterinary Drug Supervision Institute/Hubei Provincial Animal And Poultry Product Quality Supervision,Inspection and Testing Center,Wuhan,Hubei Province 430070,China;Sichuan Huahan Sanchuang Biotechnology Co.,Ltd.,Chengdu,Sichuan Province 610093,China)

机构地区：[1]中国农业科学院农业质量标准与检测技术研究所,北京100081 [2]湖北省兽药监察所/湖北省畜禽产品质量监督检验测试中心,湖北武汉430070 [3]四川华汉三创生物科技有限公司,四川成都610093

出　　处：《中国饲料》2023年第11期102-106,共5页China Feed

基　　金：中国农业科学院科技创新工程(13-IQSTAP-02);呼和浩特科技计划资助项目(2020-科技兴蒙-国创中心-9)。

摘　　要：本实验基于多重PCR与膜芯片核酸杂交技术,旨在构建同时检测饲料及原料中6种致病微生物的检测方法,并对方法的特异性、检出限及适用性进行了验证。结果表明:该方法具有良好重复性、再现性和特异性,综合检出限为培养前1~5 CFU/mL(核酸样本为0.01 ng/μL),可实现对沙门氏菌、大肠埃希氏菌O157:H7、金黄色葡萄球菌、志贺氏菌、单核细胞增生李斯特氏菌、副溶血性弧菌等6种致病微生物的高通量检测,检测结果与标准方法一致。多重PCR-膜芯片技术可成为饲料及原料中微生物污染快速、高通量筛查新的技术手段。Method for simultaneous detection of six pathogenic microorganisms in feed and feedstuffs was established based on multiplex PCR-membrane chip.The specificity,limit of detection and applicability of the developed method were verified.The method exhibited good repeatability,reproducibility and specificity.The detection limit was 1~5 CFU/mL(0.01 ng/μL for nucleic acid sample).High throughput detection of Salmonella,Escherichia coli O157:H7,Staphylococcus aureus,Shigella,Listeria monocytogenes,Vibrio parahaemolyticus was established by using PCR-membrane chip assay,and results were consistent with the standard method.Therefore,PCR-membrane chip assay could be applied for rapid and high throughput screening of microbial contamination in feed and feedstuffs.

关 键 词：多重PCR 膜芯片 致病微生物 快速检测 饲料 

分 类 号：S816.17[农业科学—饲料科学]