一种快速检测蛋鸡沙门氏菌的方法  

A rapid method for detection of Salmonellain laying hens

作　　者：李婉丽[1] 徐文龙[1,2] 李松[1] 余洋[1] 孙群[1] 

机构地区：[1]四川大学生命科学学院,成都610064 [2]四川圣迪乐村生态食品股份有限公司,绵阳621000

出　　处：《四川大学学报（自然科学版）》2015年第1期181-186,共6页Journal of Sichuan University(Natural Science Edition)

基　　金：四川省科技支撑计划(2014NZ0106);肉鸡现代产业链关键技术集成研究与产业化(2012NZ0037);国家自然科学基金(J1103518)

摘　　要：为检测蛋鸡泄殖腔拭子中沙门氏菌,在基础培养基BPW的基础上,优化出了培养仅需6 h的选择性增菌液SEM；并根据蛋鸡泄殖腔拭子中的非沙门氏菌干扰菌种类,设计了沙门氏菌特异性检测引物hilAP3.结果表明：1）低数量（1~5 CFU/50mL）沙门氏菌在优化后的一步法选择性增菌液SEM中培养6 h 后,其生长量可达10^3 CFU/mL 以上,可满足 PCR方法的检测限（10^2 CFU/mL）；2）hilAP3引物能排除蛋鸡泄殖腔拭子中的非目标菌的干扰,特异地检测其中的沙门氏菌；3）建立的SEM增菌6 h与hilAP3特异引物PCR联用的方法检测灵敏度为1~5 CFU/50mL,准确率为100%.因此,该研究中建立的优化增菌液与PCR联用法能在9h内完成沙门氏菌的检测,其灵敏度和特异性高,可应用于蛋鸡场的沙门氏菌检测.The aim of this study was to develop a method for the rapid and specific detection of Salmonel-la in the fecal swabs of laying hens.The composition of buffered peptone water （BPW）was modified in order to develop a one-step 6-hr-long culturing selective enrichment medium （SEM）,and a specific prim-er hilAP3 was designed with consideration of the interference of a variety of non-Salmonella bacteria in hen’s feces.The results showed that,after cultivation for 6 h,the growth of low quantity （1-5 CFU/50 mL）Salmonella in SEM reached 10^3 CFU/mL,which were well above the typical detection limit of PCR （1 × 10^2 CFU/mL）.Primer hilAP3 was proved to be specific for PCR amplification of Salmonella from feces of laying hens.By using this 9-hr-long method that involves one step selective culturing （6 h）and a specific primer PCR assay,as few as 1-5 CFU Salmonella per 50 mL enrichment broth could be detec-ted,and its accuracy was 100%.Hence,this optimized 9 h method is rapid,sensitive and specific,and may serve as an effective method for detection of Salmonella in fecal samples of laying hens at a large scale.

关 键 词：蛋鸡 沙门氏菌 增菌液优化 快速检测 

分 类 号：S852.61[农业科学—基础兽医学]