实时荧光定量聚合酶链反应检测血清2型猪链球菌的研究
白雪梅;张少敏;孙晖;叶长芸
【期刊名称】《疾病监测》 【年(卷),期】2011(026)003
【摘要】目的 基于TaqMan-MGB探针实时荧光定量聚合酶链反应(Real.time PCR)技术,建立针对血清2型猪链球菌的快速检测方法.方法 针对血清2型猪链球菌的csp2J基因序列,应用Beacon Designer 7.0,设计了引物和TaqMan-MGB探针,建立Real-time PCR检测方法;把目的 片段克隆到pMD18-T载体制作标准曲线;以常见致病菌及条件致病菌验证引物探针的特异性;制备血液模拟标本评价本方法在临床标本中的应用.结果 由标准曲线可知该方法可以检测到100拷贝/反应体系的目的 基因,特异性检测显示只有血清2型猪链球菌有荧光信号,其他常见致病菌及条件致病菌均无荧光信号,血液模拟标本中3.9×10<'2>cfu/ml的细菌含量可被检出.结论 以csp2J为目的 基因建立的血清2型猪链球菌Real-time PCR检测方法灵敏度高、特异性好、快速,可用于临床感染患者标本的初步筛查.%Objective To establish a rapid detection assay of Streptococcus suis serotype 2 with Taqman-MGB probe based on real-time PCR technology. Methods By using Beacon Designer 7.0, primers and Taqman-MGB probe were designed and the real-time PCR detection assay was established to detect the csp2J gene sequence of Streptococcus suis scrotype 2. The target fragment was cloned into the pMD18-T vector to make standard curve. The specificity of the probe was verified with common and conditional pathogenic bacteria, blood
实时荧光定量聚合酶链反应检测血清2型猪链球菌的研究
