猪链球菌病双重PCR诊断方法的建立

根据猪链球菌2型荚膜多糖和马链球菌兽疫亚种类M蛋白的保守区序列分别设计了2对简并引物,建立了一种能同时检测猪链球菌2型和马链球菌兽疫亚种的双重PCR方法。结果显示,该双重PCR能从100个细菌的混合纯培养物中扩增出2条目的片段。而且可以直接从病料组织中检测到相应的病原菌。用建立的双重PCR方法和细菌分离培养法平行检测人工感染的组织病料,PCR方法与细菌培养法的阳性检出率基本一致,但PCR方法的特异性好、敏感性高,简便易行,可以用于猪链球菌病的流行病学调查和实验室的快速鉴别诊断。

Establishment of multiplex PCR assay for the diagnosis of swine streptococcosis

To develop a multiplex PCR assay for the rapid and sensitive detection of Streptococcus suis type 2 and Streptococcus equi subsp.zooepidemicus in viscus from pigs,two pairs of degenerate primers were designed and synthesized according to the nucleotide sequences of conservative regions in M-like-gene of S.equi subsp.zooepidemicus and cps2J-gene of S.suis type 2.After the PCR system was optimized,two target DNAs were amplified from 100 CFU admixture bacterium of S.suis type 2 with S.equi subsp.zooe-pidemicus.The multiplex PCR assay was evaluated with tonsillar specimens from pigs infected with the two bacteria.Most of the experimental infection diagnosed by the multiplex PCR assay were confirmed by bacteriological examination.The results demonstrated that the multiplex PCR was more specific,sensitive and convenient than the bacteriological examination,and could be applied for the rapid diagnosis test and epidemiological investigation of swine streptococcosis.