猪链球菌2型的Real-time PCR定量检测研究

Quantitative detection of Streptococcus suis type 2 by Real-time TaqMan PCR

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.