沙门氏菌多重PCR检测方法的建立

Development of Multiplex PCR Detection Method for Salmonella

采用CTAB/NaCl法提取沙门氏菌及对照菌株的基因组DNA。对引物浓度、TaqDNA聚合酶用量进行优化,建立了沙门氏菌属特异基因-hut基因(495bp)、hilA基因(490bp)、invA基因(284bp)和hns基因(152bp)间的多重PCR检测方法,并进行了灵敏度测试。结果表明,建立的多重PCR检测结果与预期一致,二重PCR的检测灵敏度与单一PCR的一致,三重PCR检测灵敏度有所下降。

The genome DNAs of Salmonella and some control bacteria were extracted by CTAB/NaCl method. The multiplex PCR methods detecting synchronously among Salmonella genus special genes such as hut gene（495bp）, hilA gene（490bp）, invA gene（284bp） and hns gene（152bp） were developed by optimizing the concentration of primers and the unit of Taq DNA polymerase, and the detection limits were tested. The results showed that the detection results by multiplex PCR methods developed were same as expect, and the detection limits of the duplex PCR was same as those of single PCR, while those of triplex PCR were declined.