| 沙门氏菌通用PCR快速检测试剂盒的研制与应用 |
| |
| 引用本文： | 孔繁德,陈琼,徐淑菲,彭海滨.沙门氏菌通用PCR快速检测试剂盒的研制与应用[J].福建畜牧兽医,2007(Z1). |
| |
| 作者姓名： | 孔繁德 陈琼 徐淑菲 彭海滨 |
| |
| 作者单位： | [1]厦门出入境检验检疫局 [2]厦门市农产品质量安全检验测试中心 [3]福建农林大学动物科学学院 [4]厦门 |
| |
| 摘 要： | 根据沙门氏菌fimY基因建立了用于检测沙门氏菌的通用PCR方法。对收集的A-F群标准菌株和临床分离的60株沙门氏菌分离株和11种非沙门氏菌进行PCR检测,采用2.0%琼脂糖电泳进行检测,结果所有沙门氏菌均扩增出526bp的特异性条带,而非沙门氏菌均未扩增出任何条带。通过电泳判定结果,该法可检出扩增体系中93cfu的沙门氏菌,还可检出含3cfu的样品用BP预增菌或MM增菌后的菌液。说明该方法敏感性高、特异性强。采用直接菌体加入法、热裂解、反复冻融、CTAB碱裂解法和柱式试剂盒提取法分别提取核酸模板,结果CTAB法效果最好,但操作烦琐,而直接菌体加入法和热裂解法可以达到和柱式试剂盒同样的效果,且操作简便快速、经济、效果好,值得推广应用。
|
| 关 键 词： | 沙门氏菌 通用PCR 检测 |
Research and application on rapid detection kit of salmonella by universal PCR |
| |
| Kong Fande Chen Qiong Xu Shufei Peng Haibing.Research and application on rapid detection kit of salmonella by universal PCR[J].Fujian Journal of Animal Husbandry and Veterinary,2007(Z1). |
| |
| Authors: | Kong Fande Chen Qiong Xu Shufei Peng Haibing |
| |
| Institution: | Kong Fande1 Chen Qiong2 Xu Shufei1 Peng Haibing3 |
| |
| Abstract: | This study evaluated universal PCR to detect Salmonella species according to fimY gene.The specific primers to the fimY gene were used to amplify standard bacteria of A-F salmonella,60 salmonella separate strains and 11 non-salmonella species by PCR. Experiment results indicated that only Salmonella species generated the specific 526 bp DNA fragments on the 2.0% agarose gel and non- salmonella species controls gave negative results,demonstrating that the primers should be specific for Salmonella genus. The PCR was able to detect as little as 93 cfu Salmonella and pre-cultured(BP) or cultured(MM) bacteria liquid of the sample obtaining 3 cfu Salmonella.This indicates established PCR method is high sensitive and strongly specific.The nucleic acid is puried by five methods,including adding bacteria,hot cleavage,freezing-thaw again and again,CTAB and column reagent kit,the CTAB method IS best,but testing procedure is multifarious.Adding bacteria and hot cleavage can procure the same result as column reagent kit and have many merits including simple,rapid,economical and is worthy to generalization and application. |
| |
| Keywords: | salmonella universal PCR detection |
| 本文献已被 CNKI 维普 等数据库收录！ |
|
