摘要
目的 建立了基于TaqMan探针的Real-time PCR方法,实时、定量检测猪链球菌2型。针对GenBank公布的猪链球菌2型的csp2J基因序列设计一对引物和TaqMan探针,建立了Real-time PCR检测方法,制作标准曲线,定量检测猪链球菌2型。通过对6种细菌(9株菌株)的DNA进行扩增。结果所有4株猪链球菌2型均可产生扩增曲线,其他5株非猪链球菌2型均不产生扩增曲线。细菌纯培养物的检测敏感度为6~8CFU/PCR反应体系,相关系数均为0.99(r^2=0.99),整个试验可在1.5h内完成。建立的方法可用于猪链球菌2型的快速、定量检测。
A Real-time PCR assay was established for quantitative detection of Streptococcus suis type 2 in which the primers and probe were designed according to the csp2j gene sequence of Streptococcus suis type 2. Amplification of DNAs from 9 bacterial strains comprising 6 genera showed that all of the strains of Streptococcus suis type 2 tested(n= 4)were positive and all other species of strains tested(n=5)were negative. The sensitivity of the assay was 6-8 CFU PCR Mixture, correlation rate was 0.99 (r^2 =0.99). The assay could be completed within 1.5 h. The assays presented here can be used as a rapid screening tool for quantitative detection of Streptococcus suis type 2.
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
2006年第9期843-845,850,共4页
Chinese Journal of Zoonoses