摘要
以猪链球菌2型的荚膜多糖抗原编码基因cps2J为靶基因设计引物,利用SYBR GreenⅠ能特异地与双链DNA结合发出荧光的特性,以ABI7000为平台,建立荧光实时定量PCR检测猪链球菌2型的方法。该方法检测猪链球菌2型参考株和猪链球菌2型国内分离株都呈阳性,具有良好的特异性。整个检测过程于2h内完成,检测限度为24CFU,标准曲线的相关系数为0.997。对5份采集于猪链球菌2型病猪的病料进行检测,结果表明肝脏和脾脏最适于检测。
Streptococcus suis serotypes 2 is the most important serotype recovered from diseased pigs.This serotype is also the cause of serious infections in humans. To develop a highly rapid, specific and sensitive method of Real-time PCR assay for detection of S. suis serotype 2, a pair of primers were designed, based on the S. suis capsular polysaccharide （cps） genes specific for serotypes 2. The assay was carried out using a ABI7000 instrument and product formation was monitored continuously with the fluo-rescent double-stranded DNA binding dye SYBR Green I ,A direct correlation was determined between the fluorescence threshold cycle（Cr） and the starting quantity of S. Suis DNA （correlation coefficient 〉 0.997） .A detection limit of approximately 24 CFU of S. suis based on plate counts was determined. 5 strains of S. suis isolated from diseased pigs were positive by the Real-time PCR. Using the same Rcal-time PCR, no amplification curve were detected from S. equi group C and group D, S. suis serotype 9, ETEC O148, STEC O138, Staphylocossus aureus and Salmonella enteritidis. Of 5 samples from swine infected with S. suis serotypes 2. Both 2 liver and 2 spleen samples were positive by the Real-time PCR.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2008年第1期55-57,78,共4页
Chinese Journal of Veterinary Science
基金
国家“973”基金资助项目(2006CB504400)
