家蚕质型多角体病毒荧光定量PCR检测方法的建立  

Development of Real-time Fluorescent Quantitative PCR Method for Detecting Bombyx mori Cytoplasmic Polyhedrosis Virus

作　　者：吴萍[1,2] 钱荷英[1] 郭锡杰[1] 李龙[1,2] 陈涛[1,2] 

机构地区：[1]江苏科技大学蚕业研究所,镇江212018 [2]农业部蚕桑产业产品质量监督检验测试中心,镇江212018

出　　处：《病毒学报》2013年第1期12-16,共5页Chinese Journal of Virology

基　　金：国家自然科学基金(No.30972143);江苏省自然科学基金(BK2010353);镇江市科技支撑计划项目(NY2011028)

摘　　要：根据GenBank报道的家蚕质型多角体蛋白基因的保守序列设计特异性引物扩增118bp核苷酸片段,使用含有目的基因片段的重组质粒标准品绘制标准曲线,建立家蚕质型多角体病毒的实时荧光定量PCR检测方法。结果表明,标准曲线中模板拷贝数(X)与Ct值(Y)关系为Y=-3.582lgX+38.748,相关系数R2=0.999,构建的实时荧光定量PCR检测方法具有良好的敏感性、特异性和重复性,可用于家蚕质型多角体病毒病的快速检验及该病的流行性调查研究。A pair of specific primers were designed according to the published 118 bp conserved sequence of polyhedrin gene of Bombyx mori cytoplasmic polyhedrosis virus （BmCPV） in GenBank and a serial dilu- tions of a recombinant plasmid were prepared and used to generate standard curves,to establish a real-time fluorescent quantitative PCR method for detection of BmCPV. The results showed that the linear relation- ship between virus concentration （X） and Ct （Y） was Y=-3. 582 lgX＋38. 748 with the correlation coeffi- cient R2=0999. The method was very sensitive,specific and reproducible. It can be applied in the rapid detection of BmCPV and the prevalence investigation of this disease.

关 键 词：家蚕 质型多角体病毒 荧光定量PCR 

分 类 号：S852.65[农业科学—基础兽医学]