中国农业科学 ›› 2019, Vol. 52 ›› Issue (23): 4406-4414.doi: 10.3864/j.issn.0578-1752.2019.23.021
杨志远,段会娟,王小蕾,刘立新,赵际成,潘洁,刘月焕,林健()
收稿日期:
2019-04-23
接受日期:
2019-06-28
出版日期:
2019-12-01
发布日期:
2019-12-01
通讯作者:
林健
作者简介:
杨志远,E-mail:yangzy88@126.com
基金资助:
YANG ZhiYuan,DUAN HuiJuan,WANG XiaoLei,LIU LiXin,ZHAO JiCheng,PAN Jie,LIU YueHuan,LIN Jian()
Received:
2019-04-23
Accepted:
2019-06-28
Online:
2019-12-01
Published:
2019-12-01
Contact:
Jian LIN
摘要:
【目的】比较鸭坦布苏病毒(duck tembusu virus, DTMUV)分离株的毒力、分析E蛋白基因序列、研究抗原差异性,为鸭坦布苏病防控提供依据。【方法】将DTMUV-HB(2011)、DTMUV-AH(2014)、DTMUV-GX1(2012)、DTMUV-GX2(2015) 4株病毒接种10日龄易感鸭胚进行增殖,并利用6日龄SPF鸡胚测定ELD50。根据测定的病毒含量,将分离毒株稀释为100ELD50/0.5mL,人工感染40只180日龄健康北京鸭,进行临床症状观察、病毒分离、大体剖检,分析分离毒株的毒力有无差异。提取8株DTMUV分离病毒株的RNA,通过RT-PCR扩增E基因,分析比较不同地区的DTMUV的E基因核苷酸和氨基酸序列相似性,构建进化树。利用实验室建立的鸭坦布苏病毒血凝抑制试验分别测定4株病毒阳性血清对4株病毒的HI效价。同时采用固定病毒稀释血清法,利用C6/36细胞测定4株病毒阳性血清对4株DTMUV的血清中和效价。通过交叉血凝抑制试验和血清交叉中和试验比较4株病毒的抗原差异性(R值)。【结果】 (1)各毒株的病毒含量范围在10 4.7—10 5.3 ELD50/0.1mL。人工感染鸭试验,攻毒后3 d鸭采食量和产蛋量均显著下降,各组病毒分离阳性率均在85%以上,剖检病变主要表现为卵泡变形、出血。(2)分离株序列分析结果表明,核苷酸序列相似性为95.7%—100%,推导氨基酸序列相似性在98.2%以上。遗传进化分析表明共同构成了一个进化分支。(3)4株病毒及其阳性血清进行血凝抑制交叉试验,计算的抗原性差异R值在0.79—1.12之间;进行细胞中和交叉试验,计算的R值在0.79—1.20之间。【结论】分离的4株DTMUV在毒力、E蛋白基因和抗原性上未见显著差异。
杨志远,段会娟,王小蕾,刘立新,赵际成,潘洁,刘月焕,林健. 4株鸭坦布苏病毒的毒力、E基因序列和抗原差异性[J]. 中国农业科学, 2019, 52(23): 4406-4414.
YANG ZhiYuan,DUAN HuiJuan,WANG XiaoLei,LIU LiXin,ZHAO JiCheng,PAN Jie,LIU YueHuan,LIN Jian. Virulence, E Gene Sequence and Antigenic Difference of 4 Duck Tembusu Virus Isolations[J]. Scientia Agricultura Sinica, 2019, 52(23): 4406-4414.
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