高致病性猪繁殖与呼吸综合征(JXA1-R株)活疫苗病毒含量荧光定量PCR方法的建立

Development of qPCR method for evaluation of highly pathogenic porcine reproductive and respiratory syndrome vaccine (JXA1-R strain)

关键词:  高致病性猪繁殖与综合症  病毒含量测定  荧光定量PCR方法  

Key words:  highly pathogenic porcine reproductive and respiratory syndrome  real-time fluorescent quantitative PCR method  evaluation

摘要:

为了更加快速、准确地测得高致病性猪繁殖与呼吸综合征活疫苗的病毒含量，根据GenBank中PRRSV的保守基因序列设计合成了引物和探针，对反应条件进行优化，建立了高致病性猪繁殖与呼吸综合征（JXA1-R株）活疫苗病毒含量的荧光定量PCR方法。用该方法测定疫苗样品的病毒含量，每头份稀释100倍后，均值Ct值为17.08，病毒拷贝数均值为4.61?107copy/μL。将其与传统方法结果进行相关性分析，相关系数r=0.83，证明两种方法有很好的相关性。但是本文中只是应用该方法测定疫苗病毒含量，若要应用于疫苗效力的测定，还需要对其正确使用。

Abstract:

For the virus titre detection of highly pathogenic porcine reproductive and respiratory syndrome living vaccine , the probes and primers were designed and synthesized according to the conserved genes of PRRSV available in GenBank, and then reaction parameters were optimized to develop a real-time fluorescent quantitative PCR method for evaluation of highly pathogenic porcine reproductive and respiratory syndrome living vaccine (JXA1-R strain).Use the method to determine the amount of virus of some vaccines, average Ct value is 17.08,average concentration is 4.61×107copy/μL. According to the analysis result,the two methods are highly correlated.However, this method is only used in the determination of virus amount in this article. Its application in vaccine potency tests depends on right use of it.