新型鸭呼肠孤病毒σC蛋白遗传进化分析及多克隆抗体制备  

Genetic Evolution Analysis of Novel Duck ReovirusσC Protein and Preparation of Polyclonal Antibodies

作　　者：钟日成 林寅盛 张济培 ZHONG Richeng;LIN Yinsheng;ZHANG Jipei(Foshan Gaoming Hai Peng Aquaculture Co.,Ltd.,Foshan Guangdong 528225;Poultry Diseases Research Institute,College of Life Sciences and Engineering,Foshan University,Foshan Guangdong 528225;Wan Mu Zhou Biotechnology Co.,Ltd.,Foshan Guangdong 528225)

机构地区：[1]佛山市高明区海鹏养殖有限公司,广东佛山528225 [2]佛山大学生命科学与工程学院禽病研究所,广东佛山528225 [3]万牧洲生物科技有限公司,广东佛山528225

出　　处：《广东畜牧兽医科技》2025年第2期30-36,共7页Guangdong Journal of Animal and Veterinary Science

基　　金：佛山万牧洲生物有限公司技术开发项目(KH23279);2项专利权及水禽常见病原检测与多发病高效防控技术许可实施合同(BKH20902801)。

摘　　要：新型呼肠孤病毒(Novel duck reovirus,NDRV)是可引起番鸭、北京鸭和鹅等多种水禽,发生肝脏、脾脏不规则出血和坏死的传染性疾病,近年来广东地区水禽中该病的发病率呈上升趋势。为了解广东地区新型呼肠孤病毒遗传进化特征,该试验从广东佛山某鸭场发病番鸭中分离得到一株试验毒株,通过对该毒株σC基因序列分析,结果表明该试验毒株与NDRV同处于一个分支,并且核苷酸相似性达95.1%-99.6%;而与典型MDRV和ARV遗传距离较远,且相似性均低于50%。与SH12和DH13毒株相比,该试验毒株出现了15个氨基酸残基的变异。为了解该试验毒株σC蛋白的抗原性,进一步构建了pET32a⁃σC质粒,并建立了重组σC蛋白在BL21(DE3)内稳定表达的方法,通过Western blot鉴定该蛋白能够与NDRV阳性血清和兔多克隆抗体发生特异性结合。为进一步了解NDRV在广东地区水禽中的遗传进化特征以及基因工程疫苗的研发提供参考。Novel duck reovirus(NDRV)is capable of inducing a contagious disease in waterfowl,including ducks,Beijing ducks,and geese,which is primarily characterized by irregular hemorrhaging and necrosis of the liver and spleen.In recent years,the incidence of this disease in waterfowl in Guangdong Province has been on the rise.In order to understand the genetic evolution characteristics of the novel duck enteritis virus in Guangdong Province,a strain of NDRV was isolated from affected Muscovy ducks in a duck farm in Foshan,Guangdong.Analysis of theσC gene sequence of this strain revealed that it belongs to the same branch as NDRV,with nucleotide similarity ranging from 95.1%to 99.6%.The genetic distance from typical MDRV and ARV was far,with similarities below 50%.Compared to the SH12 and DH13 strains,this strain showed variations in 15 amino acid residues.To understand the antigenicity of theσC protein of this strain,a pET32a⁃σC plasmid was further constructed,and a method for stable expression of recombinantσC protein in BL21(DE3)was established.Western blot analysis confirmed that this protein can specifically bind to NDRV⁃positive serum and rabbit polyclonal antibodies.This study provides a reference for further understanding the genetic evolution characteristics of NDRV in waterfowl in Guangdong Province and the development of gene⁃engineered vaccines.

关 键 词：新型鸭呼肠孤病毒 σC蛋白 遗传进化分析 多克隆抗体制备 

分 类 号：S852.657[农业科学—基础兽医学]