本文采集2015年浙江省2个鸭养殖场中疑似鸭坦布苏病毒(Duck Tembusu virus,DTMUV)感染的雌麻鸭卵巢病料,经组织研磨处理,DF-1细胞纯化,成功分离了2株鸭坦布苏病毒(DTMUV),分别命名为ZJ201501和ZJ201504.利用9对两两相互重叠的PCR引物,对这2株病毒的全基因进行分段PCR扩增,经序列测定和拼接,得到病毒的全基因组序列.核苷酸序列和E蛋白氨基酸序列比对分析发现:ZJ201501和ZJ201504的核苷酸同源性高达99.9%,全长氨基酸仅有2个位点差异;与选取的19株DTMUV参考序列相比,在核苷酸水平和E蛋白氨基酸水平上,其同源性分别大于97.1%和98.4%,而ZJ201501和ZJ201504与早期分离的DTMUV FX2010遗传距离最远,这表明DTMUV在鸭群中发生了一定的变异.本研究为进一步了解DTMUV的遗传进化提供了理论依据.%Two strains ZJ201501 and ZJ201504 of Duck Tembusu virus (DTMUV) were isolated on DF-1 cells from ovary samples of shelducks suspicious of infected DTMUV and confirmed in 2015 by RT-PCR. Each of 9 overlapping segments covering the whole genomes of ZJ201501 and ZJ201504 were amplified by PCR and sequenced. The strains ZJ201501 and ZJ201504 had 99.9% nucleotide homology but had two amino acids differences. Comparing with the 19 referance strains, these two strains shared more than 97.1%nucleotide homology and more than 98.4% amino acid homology for E proteins . However, both strains showed a far genetic distance from the strain FX2010, one of the earliest DTMUV isolates, indicating that in the past years DTMUVs varied to some degree in duck populations.
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