猪溶血性链球菌的分离鉴定及溶血素基因的PCR检测

Isolation and identification of the hemolytic Streptococci from pigs and the PCR detection of the suilysin gene

为了建立简便、快捷、准确的猪溶血性链球菌实验室诊断方法,试验采用无菌操作技术将疑似猪链球菌病的猪组织接种于血平板上培养,对染色、镜检为链球菌的菌落进行分离纯化,然后进行生化鉴定、动物致病性试验、药敏试验。根据GenBank发布的猪链球菌溶血素基因(sly)全序列,设计1对特异性引物,然后抽提分离菌DNA扩增出大小为568 bp的目的片段。结果表明:使用此鉴定方法可简便、快捷、准确地检测出猪溶血性链球菌溶血素基因。

To establish a simple, fast and accurate laboratary method of testing the hemolytic Streptococci from pigs, the tissues of the piglets with suspicious disease infected by Streptococci were collected using aseptic technique and inoculated in blood agar, and it was diagnosed as Strepto coccus suis by Gram staining and microscopic observation. The colonies of Streptococci were isolated and purled, detected by the biochemical ap- praisal, animal experiment, drug sensitivity test and DNA extraction. According to the data for the sequence of Streptococcus suis hemolysin gene published in GenBank, a pair of gene specific primers were designed to amplify a fragment of568hp DNA. The result showed that the testing method was useful for simple, fast and accurate detection of hemolysin gene of the hemolytic Streptococci from pigs.