LAMP微流控芯片快速检测三种食源性致病菌  

Quick Detection of Three Food-borne Pathogens by LAMP Micro Fluidic Chip

作　　者：鞠鹤鹏[1] 戴菁 谢逸欣[1] 郑婕 陈丽璇[1] 陈洁 梁卫根 JU Hepeng;DAI Jing;XIE Yixin;ZHENG Jie;CHEN Lixuan;CHEN Jie;LIANG Weigen(Center for Disease Control and Prevention of Guangzhou Military Region,Guangzhou 510507,China)

机构地区：[1]广州军区疾病预防控制中心,广州510507 [2]广州市天河区妇幼保健院 [3]广州军区广州总医院医务部

出　　处：《解放军预防医学杂志》2018年第3期309-313,共5页Journal of Preventive Medicine of Chinese People's Liberation Army

摘　　要：建立一种结合环介导等温扩增(Loop-mediated isothermal amplification,LAMP)和微流控芯片技术检测沙门氏菌、大肠杆菌O157、金黄色葡萄球菌三种食源性致病菌的方法,实现对3种致病菌的快速检测。通过比对选取3种致病菌特异性基因、沙门氏菌侵袭蛋白A(hlyA)基因、大肠杆菌O157脂多糖编码(rfbE)基因、金黄色葡萄球菌耐药基因FemA基因作为目的基因,设计LAMP引物,通过实时LAMP筛选合格引物。设计微流控芯片,将引物冻干后固化到芯片上,制成LAMP微流控芯片。选择合适的LAMP反应体系,添加显色剂,加入芯片中,反应结果通过肉眼或仪器判读。使用标准菌株检验芯片的特异性与敏感性,并使用人工污染样品进一步检验其在实际样品检测时的敏感性。结果表明:制成的LAMP微流控芯片具有较好的特异性,3种致病菌的检测敏感性均可达到100 cfu/ml,可用于食源性致病菌的现场快速检测。Objective To establish a LAMP micro fluidic chip detection method for quickly detecting three types of foodborne pathogens： Salmonella bacteria,Staphylococcus aureus and E.coli O157. Methods The invasive protein A（ hlyA） gene of Salmonella,the lipopolysaccharide encoding（ rfbE） gene of E.coli O157,and Staphylococcus aureus drug resistance Fem A gene were selected as target genes before three pairs of primers were designed and synthesized. The appropriate primers were screened by real-time LAMP. The primers were solidified onto the designed and manufactured chip. The appropriate LAMP system was selected via experiments. The chromogenic agent was added for result interpretation by color reaction. The results were evaluated with an instrument or the naked eye. The susceptibility and specificity of the chip were tested with standard strains. The susceptibility of the chip was testified a second time with contaminated samples. Results It is showed that the LAMP chip was relatively sensitive and specific.The sensitivity for detecting the three pathogens could reach 100 cfu/ml. Conclusion This research can provide an ideal method for quick detection of food-borne pathogens.

关 键 词：环介导等温扩增 微流控芯片 食源性致病菌 快速检测 

分 类 号：TS207.4[轻工技术与工程—食品科学]