肉品中两种致病菌的双重PCR快速检测技术的研究  

Study of the Technology of Double PCR Rapid Detection of Two Species of Pathogenic Bacteria in Meat Products

作　　者：陈雪莲[1] 张晓红[1] 翟少华[1] 袁浩翔[1] 秦菊[1] 王志琴[1] 

机构地区：[1]新疆农业大学动物医学学院,乌鲁木齐830052

出　　处：《新疆农业科学》2013年第1期169-174,共6页Xinjiang Agricultural Sciences

基　　金：国家科技支撑计划课题(2011BAD47B04);新疆维吾尔自治区科技攻关项目(201130101-2);新疆维吾尔自治区重点学科基础兽医学;新疆维吾尔自治区动物生物技术重点开放实验室

摘　　要：[目的]建立一种同步检测肉品中金黄色葡萄球菌与沙门氏菌的双重聚合酶链式反应。[方法]采用7.5%氯化钠肉汤和GN增菌剂分别对金黄色葡萄球菌、沙门氏菌进行增菌,根据金黄色葡萄球菌的femA基因与沙门氏菌的invA基因设计引物,通过PCR反应对目标基因进行扩增,并对反应体系进行优化。[结果]特异性实验显示引物特异性良好,经双重体外扩增体系优化显示条带清晰,无非特异性的扩增条带。[结论]双重PCR是一种灵敏度高的快捷检测方法,可为这两种致病菌的同时检出提供新途径。[ Objective] A double PCR method for rapid and sensitive detection was constructed, which can detect Staphylococcus aureus and Salmonella simultaneously. [ Method] 7.5% sodium chloride broth and GN increasing agents were used respectively for enrichment against Staphylococcus aureus and Salmonella. In addition primers were designed in accordance with the femA gene of Staphylococcus aureus and Salmonella invA gene, and the target gene was amplified by polymerase chain reaction to filter the reaction system.[ Result] Specificity experiments showed good specificity of the primers and displayed non -specific amplified bands after the dual in vitro amplification system was optimized. [ Conclusion] The results show that the double PCR is a high - sensitivity detection method, which can provide new ways for detecting these two pathogens at the same time.

关 键 词：肉品 金黄色葡萄球菌 沙门氏菌 双重PCR检测 

分 类 号：S851.34[农业科学—预防兽医学]