养殖场环境中链球菌胞外蛋白因子PCR检测方法建立

Establishment of PCR detection method for the extracellular protein factor of Streptococcus suis in the environment around the farm

为了快速检测养殖场环境中链球菌的污染情况,试验利用链球菌胞外蛋白因子核苷酸特异序列设计引物,建立养殖场环境中链球菌的PCR快速检测方法。结果表明:从链球菌标准阳性株中获得大小为197 bp的特异性目的片段,而肠毒性大肠杆菌、沙门杆菌、金黄色葡萄球菌等均不能检出;10倍系列稀释结果显示该方法敏感性可达到单个菌株;在处理环境样品时,能检测到4×103cfu/g的环境链球菌。说明该方法具有简单、快速、准确的特点,大大缩短了检测环境中致病性猪链球菌的时间。

To rapidly detect the Streptococcus pollution in the environment around the farm, the primers were designed based＇on the unique nucleotides sequence of the extra - cellular protein factor, and then a rapid PCR detection method was established for the detection of Streptococcus in the environment. The results showed that a 197 bp fragment was obtained from the standard Gram - positive strains of Streptococcus suis, none in the Enterotoxigenie E. coli, Salmonella, Staphylococcus aureus etc. The sensitivity of this method was used for detecting a single strain in 10 - fold serial dilutions. 4 ×10^3 cfu/g Streptococcus of the environment could be detected in the treatment of environmental sample using this method. It indicates that this method has the advantages of being simple, rapid, accurate, and it can greatly shortens the time of the detection of Streptococcus suis in the environnaent.