液相基因芯片法同时检测3种食源性病原菌.pdfVIP

※分析检测 食品科学 2013, Vol.34, No.16 191 液相基因芯片法同时检测3种食源性病原菌 1 1 2 1 1 1 1 3,* 郭启新 ，张 蕾 ，张柏棋 ，赵浩军 ，韩 勤 ，毛永杨 ，彭飞进 ，李苏龙 (1.大理州质量技术监督综合检测中心，云南 大理 671000 ；2.辽宁出入境检验检疫局，辽宁 大连 116001 ； 3.黑龙江出入境检验检疫局，黑龙江 哈尔滨 150001) 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23S rDNA 、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。 关键词：液相基因芯片；金黄色葡萄球菌；单核增生李斯特氏菌；志贺氏菌 Development of Liquid Gene Chip for Simultaneous Detection of Three Species of Pathogenic Bacteria 1 1 2 1 1 1 1 3, * GUO Qi-xin ，ZHANG Lei ，ZHANG Bai-qi ，ZHAO Hao-jun ，HAN Qin ，MAO Yong-yang ，PENG Fei-jin ，LI Su-long (1. Dali Quality and Technical Comprehensive Supervision Testing Center, Dali 671000, China ；2. Liaoning Entry-Exit Inspection and Quarantine Bureau, Dalian 116001, China；3. Heilongjiang Entry-Exit Inspection and Quarantine Bureau, Harbin 150001, China) Abstract ：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 c