鼠疫耶尔森氏菌16S-23SrRNA基因间区分析

Study on 16S 23S rRNA spacer region of Y.pestis

目的 发展一种快速准确的鉴定鼠疫菌的方法。方法 Ｐ Ｃ Ｒ 反应加限制性内切酶消化的方法。结果 对来自不同疫源地的 １０３ 株鼠疫菌 １６ Ｓ２３ Ｓｒ Ｒ Ｎ Ａ 基因间区进行扩增，均可扩出两条长为 １ １４６ｂｐ 及 １ ０９０ｂｐ 的片段，扩增产物用限制性内切酶 Ｔａｑ Ｉ， Ｍ ｓｐ Ｉ消化后的酶切图谱相同。而对照菌株小肠结肠炎耶尔森氏菌，鼠伤寒沙门氏菌，大肠杆菌，痢疾杆菌，霍乱弧菌的扩增产物及酶切图谱与鼠疫菌均不相同。结论 鼠疫菌 １６ Ｓ２３ Ｓｒ Ｒ Ｎ Ａ 基因间区高度同源，基本为两种类型，长度分别为 １ １４６ｂｐ 及 １ ０９０ｂｐ，其他菌株与其明显不同；该方法将有助于快速准确的鉴定鼠疫菌。

Objective To develop a method for the indentification of Y.pestis.Methods PCR amplification and endonucleases digestion.Results The PCR amplification of 16S 23S rRNA spacer region was done on 103 Y.pestis strains that isolated from 9 different natural foci.Two fragments of 1 146bp and 1 090bp were obtained in all strains.The amplicon from each strain was digested with two endonucleases (Taq I,Msp I),and all isolates were displayed an identical pattern.Some control strains of E.coli,Y.enterocolitica,Salmonella typhimurium,Vibrio cholera,Shigella flexneri were analyzed by the approach,it was found that the amplified products and restriction patterns were different from that of Y.pestis.Conclusions The 16S 23S rRNA spacer regions are identical in all Y.pestis strains,and different from the other control strains;the approach will be helpful to the identification of Y.pestis. [