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中文摘要：
参考Gen Bank上已发表的鸭坦布苏病毒（DTMUV）NS5基因序列和产蛋下降综合征病毒（EDSV）五邻体（penton）基因序列,分别设计了检测DTMUV和EDSV的特异性引物,扩增目的片段大小分别为242 bp和181 bp。通过退火温度及引物浓度的优化,建立了针对这两种病毒的二重PCR检测方法。特异性试验结果表明,该方法可以同时检测DTMUV和EDSV,且不与鸭肝炎病毒、鸭瘟病毒、番鸭呼肠孤病毒、小鹅瘟病毒、番鸭细小病毒、H9亚型禽流感病毒、新城疫病毒以及大肠杆菌的核酸发生交叉反应;敏感性试验表明,该方法对DTMUV的最低检出限为590个拷贝,对EDSV的最低检出限为3 260个拷贝;该方法重复性良好,对不同批次的样品扩增效果良好。通过检测两种病毒感染的阳性病料各10份,结果显示,该二重PCR检测方法对阳性样品的检出率可达100%。这表明该方法可以用于鸭坦布苏病毒和产蛋下降综合征病毒的检测。
英文摘要：
Primers targeting the NS5 gene of Duck Tembusu Virus and the penton gene of Egg Drop Syndrome Virus were designed based on the data from GenBank. The two amplicons were in length of 242 and 181 basepairs. Through optimizing annealing temperature and primers concentration, the duplex PCR method was validated. This method had no cross reaction with the nucleic acid ofDHV, DPV, MDRV, GPV, MPV, H9 AW, NDV and E.coli. The nucleic acid detection limits of this method were 590 copies of DTMUVand 3260 copies of EDSV. In addition, this method had good reproducibility by three repeat tests. It showed 100% detection rate when checking the 10 positive samples. Thus, it was believed that this method could be useful for the detection of DTMUV and EDSV.
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