Abstract: Objective To develop a sensitive and specific real-time PCR method using SYBR GreenⅠ for Japanese encephalitis virus(JEV) detection. Methods Establish the optimal conditions and system of realtime PCR using SYBR GreenⅠ.Compared with the traditional RT-PCR for sensitivity and specificity.Using both assays to detect selective JEV strains,Dengue virus(Ⅰ-Ⅳ) and 90 porcine serum specimens.Results The sensitivity of this two assays,RT-PCR and real-time PCR,was no different.Compared with traditional RT-PCR,real-time PCR can save half detection time.We also proved the specificity of this new assay.Selective JEV strains were positive,while Dengue virus was negative.90 porcine serum specimens were also negative.Conclusion The results indicate that SYBR Green Ⅰ real-time PCR we established was a rapid specific and sensitive method for the detection of JEV.It can be used for investigating of JEV status.

摘要： 目的 建立一种快速、特异的SYBR GreenⅠ实时PCR方法检测流行性乙型脑炎(乙脑)病毒。方法 对SYBR GreenⅠ实时PCR检测乙脑病毒方法的反应条件、反应体系进行优化,同时与传统RT-PCR方法进行灵敏度比较,提高该方法的特异性、灵敏性。并用该方法对乙脑病毒毒株、登革病毒(Ⅰ～Ⅳ型)毒株、90份猪血清标本进行检测。结果 SYBR GreenⅠ实时PCR与传统RT-PCR检测乙脑病毒方法的灵敏度无明显区别,但检测时间缩短了1/2。该方法可检测出乙脑病毒,不能扩增登革病毒,对90份猪血清标本检测结果均为阴性。结论 建立的SYBR GreenⅠ实时PCR方法用于乙脑病毒检测具有特异、灵敏、快速等优点,可用于乙脑的病原学监测。

关键词: 流行性乙型脑炎病毒, 实时聚合酶链反应, SYBR GreenⅠ