摘要
为建立牛流行热病毒(BEFV)的荧光定量PCR检测方法,本研究根据GenBank中已登录的BEFV的G基因保守序列设计了1对特异性引物。结果显示,建立的标准曲线线性关系较好,其相关系数为0.996 3。该方法仅对BEFV检测为阳性,表明其特异性强。该方法的检测下限约为4.0×10^1 copies/μL,比普通PCR敏感性高10倍。重复性试验结果显示,该方法的组内和组间变异系数均小于0.76%。利用本方法分别对45份牛病料样品进行检测,结果检出了11份阳性样品,表明所建立的荧光定量PCR方法准确可靠,可以用于BEFV的快速诊断和定量分析。
In order to establish a real-time PCR detection method for bovine ephemeral fever virus,a pair of specific primers were designed and synthesized according to the conservative sequence of G gene in the published sequence of BEFV.The method achieved a good linear relationship with a correlation of 0.996 3.This method had no cross-reaction with the other viruses and had high specificity.And the detection sensitivity was up to 4.0×10^1 copies/μL,which is 10 times higher than the conventional PCR method.The results of repetitive experiments showed that the method had a good repeatability with coefficients of variation of intra-assay and inter-assay less than 0.76%.Using this method,the samples of 45 diseased cattle were detected,and 11 positive samples were determined.The above-mentioned results showed the detection method is accurate and reliable,and will provide technical support for the early rapid diagnosis and quantitative analysis of BEFV.
作者
楚会萌
任亚初
程凯慧
解晓莉
张亮
孙阳阳
杨宏军
CHU Hui-meng;REN Ya-chu;CHENG Kai-hui;XIE Xiao-li;ZHANG Liang;SUN Yang-yang;YANG Hong-jun(Dairy Cattle Research Center, Shandong Academy of Agricultural Sciences , Jinan 250131, China)
出处
《中国兽医科学》
CAS
CSCD
北大核心
2019年第10期1256-1260,共5页
Chinese Veterinary Science
基金
“十三五”国家重点研发计划项目(2016YFD0500904)
现代农业(奶牛)产业技术体系项目(CARS-36)
山东省农业科学院农业科技创新工程项目(CXGC2018E14)
山东省农业重大应用技术创新项目(SD2019XM006)