鼠痘病毒PCR检测方法的建立及在鼠源性生物制品检定中的应用

目的建立小鼠鼠痘病毒PCR检测方法,并应用于鼠源性生物制品的检测。方法根据鼠痘病毒基因组序列中保守的crmD片段设计一对引物,建立鼠痘病毒PCR检测方法;人工感染20只小鼠,对其肝、脾、肾、脚掌及血液采用该方法进行检测;提取鼠源性生物制品中DNA,采用该方法检测其是否存在鼠痘病毒。结果该方法可以检测到稀释至10-5感染鼠痘病毒的细胞,在BHK21细胞和牛痘病毒中均未扩增出相应片段;该方法可检测出人工感染小鼠不同组织中病毒DNA;在鼠源性生物制品中未检测出鼠痘病毒DNA。结论该方法具有良好的特异性和敏感性,可以应用于染病动物组织及鼠源性生物制品中鼠痘病毒的检测。

The Establishment of PCR Method for Mouse Poxvirus and Application in Mice-derived Biological Products Detection

Objective To establish the PCR method for mouse poxvirus（MPV） and apply it in mice-derived biological products.Method A pair of primer was designed according to the crmD gene of MPV,establish the PCR method using these primer.20 mice were infected by artificial infection,then detected the MPV DNA of liver,spleen,kidney,foot and blood of the infected mice by this method.Extract the DNA of mice-derived biological products,and detect the MPV DNA using the PCR method.Result The MPV DNA can be detected by this method when it was diluted to 10-5.And this DNA was not detected in BHK21 cell and vaccinia virus.MPV DNA can be detected in the tissue of artificial infected mice.In the mice-derived biological products,the MPV DNA was not detected.Conclusion This PCR method has nice specificity and sensibility,it can be applied in detecting of MPV infected mice and mice-derived biological products