陈睿;周跃;廖振芸;王威威;李宜海;韦平;何秀苗;
CHEN Rui;ZHOU Yue;LIAO Zhen-yun;WANG Wei-wei;LI Yi-hai;WEI Ping;HE Xiu-miao;Guangxi Key Laboratory for Polysaccharide Materials and Modifications,School of Marine Sciences and Biotechnology,Guangxi MINZU University;Aquatic Animal Husbandry and Veterinary Station of Huangsha Yaozu Township;Aquatic Animal Husbandry and Veterinary Station of Zhongyong Town;Animal Science and Technology College,Guangxi University;

• 1:广西民族大学海洋与生物技术学院广西多糖材料与改性重点实验室
• 2:桂林市临桂区黄沙瑶族乡水产畜牧兽医站
• 3:桂林市临桂区中庸镇水产畜牧兽医站
• 4:广西大学动物科学技术学院

摘要(Abstract)：

为了对某鸡群疑似传染性法氏囊病(IBD)的病例进行分子诊断、病毒分离和分子特征分析，本试验通过核酸检测调查该鸡群法氏囊组织和病毒分离物中传染性法氏囊病病毒(IBDV)的感染情况，将分离得到的毒株命名为GL1906,继而对该病毒基因组双节段的VP2高变区(vVP2)序列和VP1-b序列进行分析。结果显示，GL1906 vVP2基因的特征性氨基酸位点在222A、256I、284A、294I和299S上均符合超强毒株(vvIBDV)的特征，但279N符合致弱毒株的特征；VP1-b基因在242D、390L、393E与弱毒株一致，287A与vvIBDV一致，此外第777～782位核苷酸序列为GGTGCC,与弱毒株一致；GL1906 vVP2的核苷酸、氨基酸序列与vvIBDV的同源性最高，在系统进化树中与vvIBDV同为A3分支；GL1906 VP1-b的核苷酸、氨基酸序列与B节段属于独特来源的NN1172同源性最高，在系统进化树中同属于B3独特分支。结果表明，本试验分离株GL1906的基因组双节段vVP2和VP1-b具有不同的来源，是基因型为A3B3的基因自然重排毒株。
This study investigated an infectious bursal disease(IBD) suspected case in a chicken flock using molecular diagnostic test, virus isolation and molecular characteristic analysis. Specifically, nucleic acid test was used to detect infectious bursal disease virus(IBDV) in bursa tissue and virus culture resulted in the isolation of a strain named GL1906. Further analysis of the hypervariable region of VP2(vVP2) and VP1-b sequences in the GL1906 genome showed that the vVP2 gene contained the characteristic amino acid(aa) sites 222A, 256I, 284A, 294I and 299S in line with that of very virulent IBDV(vvIBDV), but its 279N was consistent with that of the attenuated strain; the VP1-b gene possessed the characteristic aa sites 242D, 390L and 393E in line with that of the attenuated strain, its 287A was identical to that of the vvIBDV, the nucleotide sequence at position 777-782 was GGTGCC, which was the same as that of the attenuated strain. The vVP2 gene of GL1906 showed the highest homolgy with that of the vvIBDV strains in nucleotide and amino acid level, respectively, and the phylogenetic tree analysis grouped GL1906 in the branch of A3 with vvIBDV based on vVP2. However, GL1906 shared the highest similarity in VP1-b with NN1172, a strain with B segement derived from a unique origin, and the phylogenetic tree grouped GL1906 in the branch of B3 with NN1172. Thus, the A-vVP2 and B-VP1-b of the isolate GL1906 appeared to have dirived from different sources and was a natural gene reassortant virus strain with the genotype A3B3.

关键词(KeyWords)： 传染性法氏囊病;传染性法氏囊病病毒;基因重排;分子特征
infectious bursal disease;infectious bursal disease virus;gene reassortment;molecular characteristic

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(32160824,31660717,31560706);; 广西科技重大专项(桂科AA17204057);; 广西高校中青年教师基础能力提升项目(2018KY0171);; 广西民族大学引进人才科研启动项目(2017KJQD005)

作者(Authors): 陈睿;周跃;廖振芸;王威威;李宜海;韦平;何秀苗;
CHEN Rui;ZHOU Yue;LIAO Zhen-yun;WANG Wei-wei;LI Yi-hai;WEI Ping;HE Xiu-miao;Guangxi Key Laboratory for Polysaccharide Materials and Modifications,School of Marine Sciences and Biotechnology,Guangxi MINZU University;Aquatic Animal Husbandry and Veterinary Station of Huangsha Yaozu Township;Aquatic Animal Husbandry and Veterinary Station of Zhongyong Town;Animal Science and Technology College,Guangxi University;

参考文献(References)：

• [1] Berg T P.Acute infectious bursal disease in poultry:a review[J].Avian Pathology,2000,29(3):175- 194.
• [2] 杨霖，何秀苗，禤金彩，等.一株自然基因重排鸡传染性法氏囊病病毒的分离鉴定及其分子特征分析[J].基因组学与应用生物学，2014,33 (2):282- 287.
• [3] Jackwood D J,Schat K A,Michel L O,et al.A proposed nomenclature for infectious bursal disease virus isolates[J].Avian Pathology,2018,47(6):576- 584.
• [4] Wang Y L,Fan L J,Jiang N,et al.An improved scheme for infectious bursal disease virus genotype classification based on both genome-segments A and B[J].Journal of Integrative Agriculture,2021,20(5):1372- 1381.
• [5] Rafiqul I M,Mohammed N,Tazinur R,et al.A unified genotypic classification of infectious bursal disease virus based on both genome segments[J].Avian Pathology,2021,50(2):190- 206.
• [6] Wang W W,Huang Y,Zhang Y,et al.The emerging naturally reassortant strain of IBDV (genotype A2dB3) having segment A from Chinese novel variant strain and segment B from HLJ0504-like very virulent strain showed enhanced pathogenicity to three-yellow chickens[J].Transboundary and Emerging Diseases,2021,69(4):e566- e579.
• [7] He X M,Wang W W,Chen G,et al.Serological study reveal different antigenic IBDV strains prevalent in southern China during the years 2000—2017 and also the antigenic differences between the field strains and the commonly used vaccine strains[J].Veterinary Microbiology,2019,239:108458.
• [8] He X M,Xiong Z X,Yang L,et al.Molecular epidemiology studies on partial sequences of both genome segments reveal that reassortant infectious bursal disease viruses were dominantly prevalent in southern China during 2000—2012[J].Archives of Virology,2014,159(12):3279- 3292.
• [9] 范林进，王雨龙，吴甜甜，等.我国传染性法氏囊病病毒新型变异株分析研究[J].中国预防兽医学报，2019,41(11):1164- 1169.
• [10] Kibenge F S,Jackwood D J,Mercado C C.Nucleotide sequence analysis of genome segment A of infectious bursal disease virus[J].Journal of General Virology,1990,71(3):569- 577.
• [11] 熊忠贤，何秀苗，杨霖，等.鸡传染性法氏囊病病毒流行株VP1部分序列及VP2高变区序列分析[J].中国预防兽医学报，2013,35(5):419- 422.
• [12] 韦平，龙进学，阳秀英，等.传染性法氏囊病病毒(IBDV)快速检测与分型技术[J].中国兽医学报，2004,24(4):313- 316.
• [13] 何秀苗，官丁明，韦平，等.鸡传染性法氏囊病一个自然病例的病原分离及其分子特征分析[J].中国家禽，2009,31(14):18- 22.
• [14] 何秀苗，官丁明，韦平，等.广西地区传染性囊病病毒超强毒株的分离鉴定及其VP2基因高变区的序列分析[J].中国兽医杂志，2009,45(9):7- 9.
• [15] Chen G,He X M,Yang L,et al.Antigenicity characterization of four representative natural reassortment IBDVs isolated from commercial three-yellow chickens from Southern China reveals different subtypes co-prevalent in the field[J].Veterinary Microbiology,2018,219:183- 189.
• [16] 沈巍巍，陈果，何秀苗，等.广西梧州地区近十年传染性法氏囊病病毒vVP2变化特征分析[J].动物医学进展，2017,38(3):32- 37.
• [17] 杨霖.2011—2012年间IBDV的分离鉴定及不同基因型IBDV流行株的抗原性研究[D].南宁：广西民族大学，2014.
• [18] Islam M R,Rahman S,Noor M,et al.Differentiation of infectious bursal disease virus (IBDV) genome segment B of very virulent and classical lineage by RT-PCR amplification and restriction enzyme analysis[J].Archives of Virology,2012,157(2):333- 336.
• [19] 朱向东，王永山，欧阳伟，等.传染性法氏囊病病毒自然重配株全基因组序列分析[J].中国兽医学报，2014,34(2):219- 226.
• [20] 高立，祁小乐，高玉龙，等.传染性法氏囊病病毒超强毒株HLJ-0504全基因组克隆及其新特征分析[J].中国预防兽医学报，2010,32(5):401- 404.
• [21] He X M,Chen G,Yang L,et al.Role of naturally occurring genome segment reassortment in the pathogenicity of IBDV field isolates in Three-Yellow chickens[J].Avian Pathology,2016,45(2):178- 186.
• [22] Nou?N C L,Rivallan G L,Toquin D,et al.Very virulent infectious bursal disease virus:reduced pathogenicity in a rare natural segment-B-reassorted isolate[J].Journal of General Virology,2006,87(1):2009- 2216.
• [23] 沈巍巍，陈果，何秀苗，等.反向遗传学技术在鸡传染性法氏囊病病毒研究中的应用进展[J].中国家禽，2016,38(17):45- 49.
• [24] 张建军，高巍，侯军.鸡传染性法氏囊病毒研究进展[J].中国家禽，2007,29(12):47- 49+52.
• [25] 何秀苗，官丁明，韦平，等.传染性法氏囊病病毒广西流行毒株的分离鉴定及其遗传进化分析[J].中国预防兽医学报，2009,31(8):650- 653+659.
• [26] Brandt M,Yao K,Liu M,et al.Molecular determinants of virulence,cell tropism,and pathogenic phenotype of infectious bursal disease virus[J].Journal of Virology,2001,75(24):11974- 11982.
• [27] Kasanga C J,Yamaguchi T,Munang'andu H M,et al.Genomic sequence of an infectious bursal disease virus isolate from Zambia:classical attenuated segment B reassortment in nature with existing very virulent segment A[J].Archives of Virology,2013,158(3):685- 689.
• [28] Cui P,Ma S J,Zhang Y G,et al.Genomic sequence analysis of a new reassortant infectious bursal disease virus from commercial broiler flocks in central China[J].Archives of Virology,2013,158(9):1973- 1978.
• [29] 叶丽娜，李林，沈佳，等.鸡传染性法氏囊病免疫复合物疫苗与活疫苗体外介导鸡淋巴细胞NF-κB表达的调节通路研究[J].动物医学进展，2020,41(6):42- 46.
• [30] 何秀苗，戴书剑，官丁明，等.近年来我国鸡传染性法氏囊病及其病毒分离株的变化特点[J].广西畜牧兽医，2012,28(2):67- 70.
• [31] 黄宇.广西IBDV的分离鉴定及其两种变异株全基因序列分析及致病性研究[D].南宁：广西大学，2021.