| EMA与PCR结合检测沙门氏菌方法的研究 |
| |
| 引用本文： | 赵瑜,李金磊,董鹏,狄元冉,高延玲.EMA与PCR结合检测沙门氏菌方法的研究[J].畜牧与饲料科学,2013(11):113-116. |
| |
| 作者姓名： | 赵瑜 李金磊 董鹏 狄元冉 高延玲 |
| |
| 作者单位： | [1]河南省驻马店市畜产品质量监测检验中心,河南驻马店463000 [2]河南省畜产品质量监测检验中心,河南郑州450000 |
| |
| 摘 要： | 为了建立一种快速区分样品中沙门氏菌的死细菌与活细菌的检测方法，将叠氮溴化乙锭（EMA）与PCR技术相结合。以沙门氏茵的invA为靶基因，以沙门氏菌的纯培养细胞做模板进行扩增，并进行灵敏度、特异性、曝光时间及EMA浓度试验。结果显示，灵敏度为14CFU／mL，最佳曝光时间为2min，当EMA的浓度小于18μg／mL时．EMA对活菌靶基因的扩增没有明显的抑制，而终浓度为1μg／mL的EMA，能有效抑制lxlosCFU／mL沙门氏菌死菌的扩增。EMA—PCR能有效降低沙门氏菌检测过程中的假阳性。
|
| 关 键 词： | 沙门氏菌 EMA—PCR 检测 死菌与活菌 |
Research on Detection of Salmonella Combining EMA and PCR |
| |
| Institution: | ZHAO Yu1, LI Jin-lei2, DONG Peng2, DI Yuan-ran2, GAO Yan-lin (l.Zhumadian Animal Products Quality Monitor Test Center of Henan Province, Zhumadian 463000,China;2.Henan Animal Products Quality Monitor Test Center, Zhengzhou 450000, China) |
| |
| Abstract: | In order to establish a fast detection method for distinguishing dead bacteria and live bacteria of Salmonella from samples, EMA and PCR were combined, meanwhile invA gene of Salmonella was taken as target gene, and then amplification was carried out by taking pure culture cells of Salmonella as template, furthermore sensitivity, specificity, exposure time and EMA concentration test were carried out. Results showed that： the sensitivity was 14 CFU/mL; the optimal exposure time was 2 min; when the concentration of EMA was less than 18 pLg/mL, EMA had no obvious inhibition to the amplification of live bacteria＇ target gene, while EMA at final concentration of 1 μg/mL could effectively inhibit the amplification of dead bacteria at 1 108 CFU/mL. So EMA-PCR could effectively decrease false positive rate in the detection of Salmonella. |
| |
| Keywords: | Salmonella EMA-PCR detection dead bacteria and live bacteria |
| 本文献已被 维普 等数据库收录！ |
|
