摘要
目的建立检测博尔纳病病毒(BDV)RNA的实时定量RT-PCR(real-time RT-PCR)方法。方法设计BDV-p40基因特异性引物和探针,优化反应条件;并以已知的BDV-p40基因扩增产物的克隆为标准品,建立标准曲线;然后对已知BDV RNA进行实时定量RT-PCR的特异性和敏感性检测。结果建立了特异检测BDV RNA的实时定量RT-PCR技术,并且可以检测到感染细胞中1个拷贝以上BDV RNA。结论本技术可以直接用于BDV感染的检测和定量分析。
Objective To establish the real-time RT-PCR method for detection of Boma disease vires (BDV) RNA. Methods The specific primers and the fluorescence probe of BDV-p40 were designed and synthesized; PCR products of BDV-p40 mRNA were cloned and used as standard template to develop the standard curve for samples detection;meanwhile the specificity and sensitivity of the method were performed. Results The realtime RT-PCR method for detection of BDV RNA was established and it could detect the total RNA of BDV infection cells which contained more than one copy of BDV genome. Conclusion The real-time RT-PCR method can be used to detect BDV infection and quantitatively analyse.
出处
《哈尔滨医科大学学报》
CAS
北大核心
2007年第4期293-295,共3页
Journal of Harbin Medical University
基金
国家自然科学基金资助项目(30371242)
黑龙江省杰出青年科学基金资助项目(JC04-05)