沙门氏菌简便的核酸检测技术

Detection of Salmonella by Polymerase chain Reaction

利用聚合酶链反应（Ｐｏｌｙｍｅｒａｓｅｃｈｉｎｒｅａｃｔｉｏｎ，ＰＣＲ）技术，特异性扩增沙门菌ＨＩ基因的２６９ｂｐ片段。８株标准阳性菌和３株阳性菌检测结果完全相符，并对３０株分离的疑似沙门氏菌进行了检测。采用５０μｌ体系，引物为自行设计各２０６割寡聚核苷酸，各１μＭ；ｄＮＴＰｓ各１００μＭ；Ｔａｑ酶２Ｕ。二温段水浴扩增３５循环，条件为预变性９７℃７ｍｉｎ，变性９４℃６０ｓ，复性和延伸６０℃６０ｓ。

HI priners which targeted a 269 bp region of HI gene inSalmonella,were used in the polymerase chain reaction(PCR). The standard PCR was done i 50 μl volume. It contained 1μM of eachrinier,2.5mM Mgcl 2,200μM dNTPs,104 ～ 108 copies of templates DNA. The amplification was performed by mannual operation between twotemperature control scations . Temperature cycle was designde in former denaturation at 97℃ 7min,35 cycles with denaturation 94℃ 60 s anneating and extension at 60℃ 60s,and last estension at 72℃ 7min.