猪链球菌2型及其毒力因子检测多重PCR的建立与应用

Multiplex PCR Assays for Simultaneous Detection of Serotypes 2 and Two Virulence Factors(mrp/epf) of Streptococcus suis

根据相关文献设计并合成引物,建立了能同时检测猪链球菌2型(cps)及其重要毒力因子溶菌酶释放蛋白(mrp)和细胞外因子(epf)的多重PCR方法。目的片段的大小分别为885bp(mrp)、675bp(cps)和443bp(epf)。对参考菌株、人工攻毒病料和临床收集病料的检测结果显示,该多重PCR特异性强、敏感性高,可直接从临床病料中检测出猪链球菌2型,并能鉴定其毒力因子表型。

A multiplex PCR was developed for the rapid and sensitive detection of Streptococcus suis serotype 2 （SS2） strains and two virulence factors genes：muramidase-released protein（mrp） and extracellular factor（epf） in viscus from pigs. The PCR products of the mrp,epf and cps were 885 bp,443 bp and 675 bp respectively. When DNA from the bacteria of S. agalactiae,P. pneurnotropica and S. eqiusimilis were used as templates for amplification,no product was detected. All samples from artificially infected swine and rabbits were positive by multiplex PCR. Of the 34 clinical specimens,8 specimens were tested positive by the bacteriological examination,but 18 specimens were positive by multiplex PCR assays. The results indicated that the multiplex PCR was more sensitive than the bacteriological examination.