SYBR Green II荧光定量PCR结合熔解曲线鉴别不同亚型兔病毒性出血症病毒方法的建立  

DIFFERENTIATION OF SUBTYPES OF RABBIT HEMORRGAGIC DISEASE VIRUS USING SYBR GREEN II REAL-TIME PCR AND DIFFERENT MELTING CURVES

作　　者：谭永贵 刘腾[1] 朱杰[1] 郭慧敏[1] 吴巧梅[1] 李琦[1] 缪秋红[1] 陈宗艳[1] 李传峰[1] 刘光清[1] TAN Yong-gui;LIU Teng;ZHU Jie;GUO Hui-min;WU Qiao-mei;LI Qi;MIAO Qiu-hong;CHEN Zong-yan;LI CHUAN-feng;LIU Guang-qing(Shanghai Veterinary Research Institute, CAAS, Shanghai 200241, China)

机构地区：[1]中国农业科学院上海兽医研究所,上海200241

出　　处：《中国动物传染病学报》2017年第1期7-11,共5页Chinese Journal of Animal Infectious Diseases

基　　金：国家重点研发计划项目(2016YFD0500108);国家自然科学基金项目(31502068);上海市科技兴农重点攻关项目

摘　　要：本研究建立了一种荧光定量PCR(Real-time fluorescence quantitative PCR)结合熔解曲线区分不同亚型兔病毒性出血症病毒(Rabbit hemorrhagic disease virus,RHDV)方法。根据GenBank数据库中已公布的不同亚型RHDV(经典RHDV和RHDV2)的衣壳蛋白VP60序列保守区设计了1对特异性引物,利用SYBR Green II荧光染料,在实时荧光定量PCR方法的基础上结合熔解曲线进行分析,经典RHDV和RHDV2熔解温度(Tm)分别为(86.3±0.1)℃和(85.1±0.1)℃,扩增产物的熔解曲线分析均只出现单特异峰。结果表明,本研究建立的方法能够快速地鉴别经典RHDV和RHDV2。The real-time fl uorescence PCR in combination of unique melting curves was developed to distinguish Rabbit hemorrhagic disease virus(RHDV)and RHDV2.A pair of specifi c primers were designed according to the published GenBank database of RHDV and RHDV2capsid protein VP60and labeled with SYBR Green fl uorescent dye II.The melting temperatures of RHDV and RHDV2were(86.3+0.1)℃and(85.1-0.1)℃,respectively,and single specifi c peaks were observed.These results indicated that the method developed here was able to quickly identify RHDV and RHDV2.

关 键 词：兔病毒性出血症病毒 荧光定量PCR 熔解曲线 鉴别 

分 类 号：S852.659.6[农业科学—基础兽医学]