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.
作者
魏书林
李文静
宋荣
李丽蓓
王培龙
冯超林
张维
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)
出处
《中国饲料》
北大核心
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)。
关键词
多重PCR
膜芯片
致病微生物
快速检测
饲料
multiplex PCR
membrane chips
pathogenic microorganism
rapid detection
feed
