一株鸽副黏病毒Ⅰ型分离鉴定及致病性分析  

Isolation,Identification and Pathogenicity Analysis of a Pigeon Paramyxovirus-1 Strain

作　　者：张姗 刘大虎 刘宝京 梁琳[1,2] 梁瑞英 汤新明 仇旭升 丁铲[5] 丁家波[1,2] 侯绍华[1] ZHANG Shan;LIU Dahu;LIU Baojing;LIANG Lin;LIANG Ruiying;TANG Xinming;QIU Xusheng;DING Chan;DING Jiabo;HOU Shaohua(Institute of Animal Science,Chinese Academy of Agricultural Sciences,Beijing 100193,China;Key Laboratory of Animal Biosafety Risk Prevention and Control(North)/Key Laboratory of Veterinary Biological Products and Chemicals,Ministry of Agriculture and Rural Affairs,Beijing 100193,China;Tianjin Key Laboratory of Agricultural Animal Breeding and Healthy Husbandry,College of Animal Science and Veterinary Medicine,Tianjin Agricultural University,Tianjin 300392,China;Bejing Xinhexiang Technology Co.,LLC,Beijing 100085,China;Shanghai Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Shanghai 200241,China)

机构地区：[1]中国农业科学院北京畜牧兽医研究所,北京100193 [2]农业农村部动物生物安全风险预警及防控重点实验室(北方)/农业农村部兽用生物制品与化学药品重点实验室,北京100193 [3]天津农学院动物科学与动物医学学院,天津市农业动物繁育与健康养殖重点实验室,天津300392 [4]北京信和翔科技有限责任公司,北京100085 [5]中国农业科学院上海兽医研究所,上海200241

出　　处：《畜牧兽医学报》2024年第9期4051-4060,共10页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：上海市肉鸽产业技术体系建设(沪农科产字(2023)第12号);中国农业科学院创新工程计划(ASTIP-IAS15)。

摘　　要：在天津的病死鸽病料中分离鉴定鸽副黏病毒Ⅰ型(pigeon paramyxovirus-1,PPMV-1),并对其进行遗传演化、生物学特性、致病性分析,为鸽副黏病毒Ⅰ型疫苗的研发提供科学依据。采用RT-PCR方法进行病原检测,在SPF鸡胚上分离纯化病毒;经遗传演化分析、致病性分析及动物回归试验对病毒分离株进行全面分析。结果显示:病死鸽的脏器样品经RT-PCR鉴定为NDV核酸阳性;鸡胚分离纯化后得到1株鸽副黏病毒Ⅰ型毒株,命名为Pigeon/TJ/CH/020/2020(TJ20)。通过对病毒F基因测序及进化树分析,确定该分离毒株属于ClassⅡ类Ⅵ.2.1.1.2.2(原Ⅵk)基因亚型,裂解位点的氨基酸残基为112 R-R-Q-K-R-F^(117),符合NDV强毒株的分子特征。交叉血凝抑制试验显示TJ20株与LaSota疫苗株存在明显的抗原性差异。TJ20株的鸡胚半数感染量(EID_(50))为10^(-8.38)·0.1 mL^(-1)、细胞半数感染量(TCID_(50))为10^(-8.64)·0.1 mL^(-1)、鸡胚平均致死时间(MDT)为62 h、1日龄雏鸡脑内接种致病指数(ICPI)为1.19。动物感染试验结果表明,1月龄鸽人工感染TJ20株后第4天开始出现嗜睡、垂翅、腹泻、瘫痪、斜颈扭头等新城疫典型临床症状,发病率和死亡率均为100%。本研究从病死鸽组织中成功分离出一株具有高致病性的Ⅵ.2.1.1.2.2(原Ⅵk)基因亚型的TJ20毒株,为后续鸽副黏病毒Ⅰ型疫苗研发提供了重要的生物学材料。A strain of pigeon paramyxovirus-1(PPMV-1)was isolated from the dead pigeon tissue in Tianjin region.To provide scientific basis for the development of PPMV-1 vaccines,the genetic evolution,biological characteristics,and pathogenicity of the virus were analyzed.The pathogens were identified by RT-PCR,virus isolation,purification by chicken embryos,genetic evolution analysis,pathogenicity analysis,and animal regression tests.A strain of PPMV-1 was isolated from the dead pigeons and named Pigeon/TJ/CH/020/2020(TJ20).Sequence analysis and homology analysis of the virus F gene showed that the strain belonged to sub-genotypeⅥ.2.1.1.2.2(Ⅵk),and the amino acid residues of the cleavage site were 112 R-R-Q-K-R-F^(117),which were consistent with the molecular characteristics of NDV virulent strains.Hemagglutination inhibition revealed a noticeable difference between TJ20 and LaSota.The values of TCID_(50),EID_(50),MDT and ICPI were 10^(-8.64)·0.1 mL^(-1),10^(-8.38)·0.1 mL^(-1),62 h,1.19.The animal infection experiment found that pigeons showed clinical symptoms on 4 dpc(day 4 post-challenge).The incidence rate and mortality were 100%,and the cloacal detoxification rate could reach 100%on 5 dpc.TJ20 was highly pathogenic to pigeons.This study successfully isolated a highly pathogenic sub-genotypeⅥ.2.1.1.2.2(Ⅵk)TJ20 strain from samples of dead pigeons,providing important biological materials for the research and development of vaccine of PPMV-1 in the future.

关 键 词：鸽副黏病毒Ⅰ型 分离鉴定 Ⅵ.2.1.1.2.2(原Ⅵk)基因亚型 致病性 

分 类 号：S852.659.5[农业科学—基础兽医学]