尼帕病毒Taqman qRT-PCR检测方法的建立

Establishment of Taqman qRT-PCR Method for Detection of Nipah Virus

尼帕病毒(NiV)是一种高致病性人畜共患病原体,可引起人类致死性脑炎和严重的呼吸系统疾病,目前尚无有效的抗NiV药物和疫苗,建立NiV检测方法十分必要。建立针对尼帕病毒N基因的Taqman qRT-PCR检测方法,应用于样本尼帕病毒检测及定量。针对尼帕病毒N基因保守区域设计引物和探针,建立Taqman qRT-PCR检测方法,评价该检测方法灵敏度、特异性和可重复性,并应用此方法进行样本检测。本研究建立的Taqman qRT-PCR检测方法可以特异性的检测尼帕病毒,检测灵敏度为10~2拷贝/μL,标准曲线线性范围为1.0×10~9~1.0×10~2拷贝/μL,可重复性较好,能够有效检出尼帕病毒马来西亚病毒株和孟加拉病毒株,可用于样本尼帕病毒的检测及定量。

A Taqman qRT-PCR method for detection of Nipah virus(NiV)N gene was established and applied to the detection and quantification of NiV samples. Primers and probe were designed based on the conserved region of NiV N gene,and a Taqman qRT-PCR detection method was established. The sensitivity,specificity,and repeatability of the method were analyzed,and the method was used for sample detection. The analytical detection limit of this Taqman qRT-PCR method established in this study was 10~2 copies/μL and allowed quantitation ranging from 1.0×10~9 to 1.0×10~2 copies/μL. This method showed good specificity and repeatability. NiV Malaysia strain and Bangladesh strain could be effectively detected by the Taqman qRT-PCR,so it can be used for the detection and quantification of NiV samples.