中国动物传染病学报Issue(4)：46-50,5.
鸭坦布苏病毒实时荧光定量RT-PCR检测方法的建立及应用
DEVELOPMENT AND APPLICATION OF REAL-TIME RT-PCR FOR DETECTION OF DUCK TEMBUSU VIRUS
靳宇田 1黄显明 1许秀梅 1尹秀凤 1徐延伟 1朱鹏 1张小飞1
作者信息
- 1. 南京天邦生物科技有限公司生物技术研究所，南京211102
- 折叠
摘要
Abstract
A pair of primers and a TaqMan probe were designed according to the reported genomic sequence of Duck Tembusu virus (DTMUV) (GenBank accession No. JX273153) and an accurate and rapid real-time RT-PCR assay was then developed. The real-time RT-PCR method specifically detected DTMUV and had no cross-reaction with of other duck pathogens. The sensitivity of the assay was 25copies/μL. The linear correlation was observed at DNA concentrations from 2.5×101copies/μL to 2.5×107copies/μL. The equation of the assay was Ct=–3.323×Lg (conc)+43.70. The correlation coefficient was 0.997 and PCR efficiency reached up to 100%. Twenty-nine samples were collected from diseased ducks on different farms in Jiangsu and Anhui provinces and detected using this method. As a result, 27 samples were tested positive (93.10%). The real-time RT-PCR assay developed in the present study was a rapid, sensitive and specific method for reliable and quantitative detection of DTMUV, which can be used for clinical diagnostics and molecular epidemiology.关键词
鸭坦布苏病毒/荧光定量RT-PCR/应用Key words
Duck Tembusu virus (DTMUV)/TaqMan real-time RT-PCR/application分类
农业科技引用本文复制引用
靳宇田,黄显明,许秀梅,尹秀凤,徐延伟,朱鹏,张小飞..鸭坦布苏病毒实时荧光定量RT-PCR检测方法的建立及应用[J].中国动物传染病学报,2013,(4):46-50,5.
