摘要
目的:建立一种能同时检测3种食源性病原菌的液相基因芯片方法。方法:针对金黄色葡萄球菌23SrDNA、志贺氏菌iapH、单核增生李斯特氏菌iap基因分别设计引物和探针,通过多重聚合酶链式反应扩增获得大小为246、112bp和174bp的目的片段;将探针偶联到编码微球上,制成基因芯片,与目的片段进行杂交,建立3种食源性病原菌的液相基因芯片方法;并评价该方法的特异性、灵敏度。结果:建立的液相基因芯片检测方法特异性好、灵敏度高。对金黄色葡萄球菌、单核增生李斯特氏菌、志贺氏菌的检测灵敏度分别为38、44、21CFU/mL。结论:成功建立了3种致病菌的液相基因芯片检测方法。
Objective： To establish a method for simultaneous detection of three species of foodborne pathogenic bacteria using liquid gene chips.Method： Three sets of specific primers and probes were designed according to the gene 23S rDNA of Staphylococcus aureus,the gene iap of Listeria monocytogenes and the gene ipaH of Shigella spp.The multiple PCR reaction systems gave rise to target fragments of 246,112 bp and 174 bp,respectively.The target fragments were captured by microspheres coupled with the gene-specific probes,finally establishing liquid gene chips.Results： The method was highly sensitive and specific.The sensitivity for Staphylococcusaureus,Listeria monocytogenes and Shigella spp.were 38,44 CFU/mL and 21 CFU/mL,respectively.Conclusion： We here report a novel detection assay using liquid gene chips for three foodborne pathogenic bacteria,which provides a highthroughput system for rapid detection of foodborne pathogenic bacteria.
出处
《食品科学》
EI
CAS
CSCD
北大核心
2013年第16期191-195,共5页
Food Science
基金
国家质检总局科技计划项目(2009IK161)
