隐孢子虫种株的RAPD分析

Random Amplified Polymorphic DNA Analysis of Cryptosporidium Species and Strains

目的 对鼠隐孢子虫 (Cryptosporidiummuris)、小球隐孢子虫 (Cryptosporidiumparvum)和火鸡隐孢子虫 (Cryptosporidiummeleagridis) 3个种 8个虫株进行随机扩增多态性DNA (RAPD)分析。 方法 从2 0 0个引物中选用重复性较好的 2 6个引物对所有虫株进行随机扩增。结果 RAPD分析结果表明 :①在隐孢子虫不同种 (C muris、C parvum和C meleagridis)之间的基因存在明显的差异 ,平均遗传距离为 7 0 37;②同种不同分离株之间均有一定差异 ,平均遗传距离为 5 16 2。结论 RAPD技术能鉴别 8个隐孢子虫分离株共有和各自特有基因片段。

Objective To analyse the genetic heterogeneity of 8 isolates of Cryptosporidium belonging to three species ( Cryptosporiduim muris,Cryptosporiduim parvum and Cryptosporidium meleagridis ) using RAPD technique.Methods DNA extracts of the 8 isolates were amplified by PCR using 26 out of 200 primers. The amplified products were separated by agar electrophoresis followed by staining with ethidium bromide.Results RAPD analysis revealed:① Obvious differences among species were found, the average genetic distance being 7\^037;② Certain differences among different isolates of the same species were found,the average genetic distance being 5\^162.Conclusion RAPD could differentiate the common and respective gene fragments of the 8 isolates of Cryptosporidium .