鸭坦布苏病毒一步法RT-PCR检测方法的建立和应用

摘要

Duck Tembusu virus ( DTMUV) is a new flavivirus that can cause Duck hemorrhagic ovaritis, a duck infectious disease with egg-drop. It is important to establish a pathogen detection assay for diagnosis and epidemiologi-cal studies. A pair of primers was designed based on the analysis of TMUV sequences. The one-step RT-PCR method had been successfully developed for detection of DTMUV through optimization of viral RNA extraction and RT-PCR assay. The results showed that 1. 82 TCID50 of virus could be detected in two hours. The rate of positivity in 106 clinically suspected tissues and organs of diseased duck subjected to detection by RT-PCR was 46. 2%. The results indicated that DTMUV is highly prevalent in Zhejiang and its neighboring provinces.%鸭坦布苏病毒(Duck Tembusu virus,DTMUV)是一种新发现可引起鸭产蛋下降的新型黄病毒,建立一种病原学检测方法对于疫病的诊断和流行病学研究具有重要意义.在对DTMUV序列的比对分析的基础上,设计了1对特异性引物,通过优化PCR反应条件,建立了DTMUV的一步法RT-PCR检测方法.该方法具有特异、快速、敏感、准确等特点,可以检出1.82个TCID50的病毒核酸.对2010年至2011年6月份前的106份产蛋下降鸭群临床病料进行了检测,结果显示样品阳性率为46.2％,表明DTMUV在浙江及其周边省份的产蛋鸭群具有较高的感染率.