一种快速检测蛋鸡沙门氏菌的方法
A rapid method for detection of Salmonella in laying hens
作者：李婉丽(四川大学生命科学学院)；徐文龙(四川大学生命科学学院)；李松(四川大学生命科学学院)；余洋(四川大学生命科学学院)；孙群(四川大学生命科学学院)
Author：LI Wan-Li(College of Life Sciences, Sichuan University)；XU Wen-Long(College of Life Sciences, Sichuan University)；LI Song(College of Life Sciences, Sichuan University)；YU Yang(College of Life Sciences, Sichuan University)；SUN Qun(College of Life Sciences, Sichuan University)
收稿日期：2013-08-29 年卷（期）页码：2015,52(1):181-186
期刊名称：四川大学学报: 自然科学版
Journal Name：Journal of Sichuan University (Natural Science Edition)
关键字：蛋鸡; 沙门氏菌; 增菌液优化; 快速检测
Key words：Laying hens; Salmonella; Enrichment medium optimization; Rapid detection
基金项目：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)
中文摘要
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103 CFU/mL以上, 可满足PCR方法的检测限(102 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9 h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.
英文摘要
The aim of this study was to develop a method for the rapid and specific detection of Salmonella 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 primer 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 103 CFU/mL, which were well above the typical detection limit of PCR (1 × 102 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 detected, 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.
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