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

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.
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 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 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