摘要
目的建立2个分开的多重聚合酶链反应(PCR)体系,以及对猪链球菌7种主要毒力因子的检测。方法根据猪链球菌7种主要毒力因子mrp、epf、sly、gdh、gapdh、orf2和fbps的基因序列,设计和合成7对特异性引物,通过对它们在2个分开反应体系PCRⅠ和PCRⅡ中的组合和优化,建立猪链球菌7种主要毒力因子的2组多重PCR检测方法,并对实验室的29株背景明确的猪链球菌保存菌株进行检测。结果29株猪链球菌菌株的检测结果和菌株的背景情况一致,阳性、阴性对照均成立。结论此猪链球菌7种主要毒力因子的2组分开体系的多重PCR检测方法,特异性和敏感性均好。
Objective To develop a rapid and sensitive assay system to detect the seven virulence factors of Streptococcus suis,using two seqarate multiplex PCR assays. Methods The seven virulence factors were detected based on the gene sequences of mrp, epf, sly gdh, gapdh, orf2 and fbps. PCR primer pairs were produced for each of the 7 factors and the assay method, was developed and optimized. These genes were tested in 29 bacterial isolates by the multiplex assay. Results All PCR products were analyzed by electrophoresis on 2% agarose gels. Detection of all 29 Streptococcus su/s strains was confirmed by bacteriological examination. The assay was validated by the positive and negative controls. Conclusion The results demonstrate that the multiplex PCR assay is a highly specific and sensitive diagnostic tool for the detection of seven virulence factors of Streptococcus suis.
出处
《微生物与感染》
2007年第1期30-33,共4页
Journal of Microbes and Infections
关键词
猪链球菌
毒力因子
多重聚合酶链反应
Streptococcus suis
Virulence-associated factors
Multiplex polymerase chain reaction
