摘要
为建立灵敏、高效的登革病毒检测方法,本试验拟以包含登革病毒3′端保守序列的质粒pUC57-DENV为模板,建立检测DENV的荧光定量PCR方法。结果显示,该检测方法在4.88×10^1~4.88×10^9copies/μL范围内呈良好的线性关系,相关系数为0.999,斜率为-3.653。最低检测限度为4.88 copies/μL,对照组未出现特异性扩增,所有稀释度的标准品在80.20℃出现特异性熔解峰。组内和组间试验的变异系数均小于1%。本试验建立的SYBR Green Ⅰ荧光定量PCR方法,可为登革病毒早期检测提供技术支持。
To establish a sensitive and efficient detection method for dengue virus.This experiment intends to use the plasmid pUC57-DENV containing the 3’-end conserved sequence of dengue virus as a standard template to establish fluorescence quantitative PCR method for detecting DENV.The results showed a good linear relationship between 4.88 ×10^1-4.88 ×10^9 copies/μL,and the correlation was 0.999 with a slope of-3.653.The lowest detected limit was 4.88 copies/μL,and there was no specific amplification in the control group.Standard templates in various dilutions all showed specific melting peaks at 80.20 ℃.The coefficient of variation for both within and between trials was less than 1%.This experiment established the SYBR Green Ⅰ real-time PCR method for dengue virus,which can provide technical support for the early detection of dengue virus.
作者
汪伟
于宁
刘宇梦
李成辉
韩知晓
杜倩
孙文超
鲁会军
金宁一
WANG Wei;YU Ning;LIU Yu-meng;LI Cheng-hui;HAN Zhi-xiao;DU Qian;SUN Wen-chao;LU Hui-jun;JIN Ning-yi(College of Animal Science and Technology,Guangxi University,Nanning 530001,China;Institute of Military Veterinary,The Academy of Military Medical Sciences,Changchun 130122,China;Agricultural College,Yanbian University,Yanji 133000,China;Institute of Virology,Wenzhou University,Wenzhou 325035,China)
出处
《中国兽医科学》
CAS
CSCD
北大核心
2020年第11期1353-1359,共7页
Chinese Veterinary Science
基金
广西自然科学基金项目(2012GXNSFAA053073)
浙江省青年基金项目(LQ19C180001)
温州市基础性科研项目(N20190005,N20180010)。
