鸭源新型鹅细小病毒的分离鉴定与VP1基因序列分析  

Isolation and Identification of a Novel Goose Parvovirus from Duck and Sequence Analysis on VP1 Gene

作　　者：刘博[1,2] 祖立闯 魏中锋 谷英华 毕研丽 王文秀[1,5] LIU Bo;ZU Lichuang;WEI Zhongfeng;GU Yinghua;BI Yanli;WANG Wenxiu(Shandong Binzhou Animal Science&Veterinary Medicine Institute,Binzhou,Shandong 256600;Shandong Lvdu Biotechnology Co.,Ltd.,Binzhou,Shandong 256600;Zibo Animal Disease Prevention and Control Center,Zibo,Shandong 255000;Heze Animal Disease Prevention and Control Center,Heze,Shandong 274006;Shandong Academician Workstation,Binzhou,Shandong 256600)

机构地区：[1]山东省滨州畜牧兽医研究院,山东滨州256600 [2]山东绿都生物科技有限公司,山东滨州256600 [3]淄博市动物疫病预防与控制中心,山东淄博255000 [4]菏泽市动物疫病预防控制中心,山东菏泽274006 [5]山东省院士工作站,山东滨州256600

出　　处：《中国家禽》2022年第12期34-39,共6页China Poultry

基　　金：山东省外专双百计划项目(WST2018014);2021年海南省农业科学院院级项目重点实验室开放课题(202106)

摘　　要：为确定发病鸭场的致病原,掌握该致病原的遗传进化特征,研究从山东省某鸭场采集到1份短喙与矮小综合征(SBDS)疑似病例病料,采用永生化鹅胚上皮细胞系(GEEC)对病料进行病毒分离培养,观察细胞病变,测定分离毒株的血凝性、病毒滴度(TCID_(50))、对鸭胚的致病性,并进行VP1基因序列比对及相似性分析。结果显示:临床病料分离培养的病毒可引起GEEC出现细胞明显皱缩、聚集、折光性增强等典型细胞病变,表明分离到一株鸭源NGPV毒株,命名为SDHZ株,该病毒对1%鸡红细胞无凝集性,TCID_(50)高达10^(-6.25)/mL,鸭胚半数致死量(ELD_(50))为10^(-4.5)/mL,接种鸭胚死亡出现全身及肝脏出血、短喙、发育迟缓等典型症状,VP1基因序列与樱桃谷鸭中分离的新型鹅细小病毒(NGPV)相似性为95.3%~98.0%,与GPV经典毒株的相似性为89.3%~94.6%。表明该鸭场本次疫病病原为NGPV,为进一步研究NGPV致病机制及制定综合防控措施提供参考。To determine the pathogen of the duck farm and master the genetic evolution characteristics of the pathogen, a sample from suspected short beak and short stature syndrome(SBDS) was collected from a duck farm in Shandong province. The virus was isolated and cultured on immortalized goose embryo epithelial cell line(GEEC), which displayed cytopathic effect. The hemagglutination, virus titer(TCID_(50)) and pathogenicity to duck embryos were measured, and the VP1 gene sequence alignment and similarity analyses were carried out. The results showed that clinical samples could cause GEEC to have typical cytopathy such as obvious cell shrinkage, aggregation and enhanced refraction. Sequencing analysis showed that a duck derived NGPV virus strain was isolated from the disease material and named SDHZ, which showed no agglutination with 1% chicken erythrocytes;The TCID_(50) of SDHZ strain was as high as 10^(-6.25)/mL;The median lethal dose(ELD_(50)) of the isolated virus SDHZ strain to duck embryos was 10^(-4.5)/mL and the dead embryos showed typical symptoms such as systemic and liver bleeding, short beak, growth retardation;The similarity of VP1 gene sequence with the new goose parvovirus(NGPV) isolated from cherry valley duck was 95.3% to 98.0%, and with the classic GPV strain was 89.3% to 94.6%. In summary, the pathogen of the duck intestinal disease was NGPV,it provided a reference for further studying the pathogenic mechanism of NGPV and formulating comprehensive prevention and control measures.

关 键 词：新型鹅细小病毒 永生化鹅胚上皮细胞系 分离鉴定 VP1基因 序列分析 

分 类 号：S852.65[农业科学—基础兽医学]