牛流行热病毒云南流行毒株G基因遗传多样性分析  

Analysis of the Genetic Diversity of Bovine ephemeral fever virus G Gene in Yunnan Province of China

作　　者：彭海芬 王国君 段新慧[3] 刘丽仙[4] 杨莎莎 张以芳 李文贵 PENG Haifen;WANG Guojun;DUAN Xinhui;LIU Lixian;YANG Shasha;ZHANG Yifang;LI Wengui(College of Veterinary Medicine,Yunnan Agricultural University,Kunming 650201,China;Lancang Vocational High Schools,Lancang 665600,China;Faculty of Animal Science and Technology,Yunnan Agricultural University,Kunming 650201,China;Yunnan Vocational College of Agriculture,Kunming 650212,China)

机构地区：[1]云南农业大学动物医学院,云南昆明650201 [2]澜沧县职业高级中学,云南澜沧665600 [3]云南农业大学动物科学技术院,云南昆明650201 [4]云南农业职业技术学院,云南昆明650212

出　　处：《华北农学报》2024年第S1期281-288,共8页Acta Agriculturae Boreali-Sinica

基　　金：云南省科技厅创新引导与科技型企业培育计划(202204BI090001)。

摘　　要：为调查云南大理州一处奶牛养殖小区BEFV的感染情况及其流行毒株G基因的遗传多样性。对该大理州奶牛养殖小区的10份疑似病例,参考GenBank中牛流行热病毒的G基因序列,设计并合成2对特异性扩增G基因,测序并完成序列分析。同时,采60份血清样本进行抗体检测。结果显示,对10份疑似BEF病牛全血样的检测结果,6份呈BEFV核酸阳性。去除重复序列,获得4株牛流行热病毒云南流行株G基因扩增、测序,序列全长均为1872 bp,编码623 aa。4株毒株之间组内核苷酸序列相似性为99.7%~99.9%,遗传进化分析与亚洲毒株聚为一类,并独自为一个细小分支。与泰国2017年分离TH-NP0065毒株和中国的毒株关系最近。与我国疫苗株JB76H的氨基酸同源性比较,发现7个氨基酸突变位点,为K^(53)→N^(53)、K^(62)→R^(62)、P^(183)→S^(183)、G^(263)→E^(263)、K^(461)→E^(461)、K^(457)→R^(457)、I^(475)→M^(475)。除中和抗原位点G1外,G2、G3、G4均出现变异位点。抗体检测试剂盒检测结果,抗体浓度≥52.5 ng/L临界值的样品有19份,阳性率为31.67%。综上所述,该奶牛养殖小区存在BEFV的感染,且流行株G基因存在多位点变异。To investigate the infection of BEFV and the genetic diversity of its prevalent strain G gene in a dairy farming community in Dali Prefecture,Yunnan Province.It designed and synthesized two pairs of specific amplified G genes for 10 suspected cases in dairy cattle breeding community of Dali Prefecture,with reference to the G gene sequence of bovine ephemeral fever virus in GenBank,sequenced and completed sequence analysis.At the same time,60 serum samples were collected for antibody detection.The results showed that 6 out of 10 whole blood samples of cattle suspected of BEF were positive for BEFV nucleic acid.The G gene of four bovine ephemeral fever virus Yunnan epidemic strains was amplified and sequenced by removing the repetitive sequence.The total sequence length was 1872 bp,encoding^(62)3 aa.The nucleotide sequence homology among the four strains was 99.7%-99.9%.Phylogenetic analysis showed that the four strains were grouped together with Asian strains and were a small branch alone.It had the closest relationship with TH-NP0065 strain isolated from Thailand in 2017 and Chinese strain.Compared with the amino acid homology of the vaccine strain JB76H in China,seven amino acid mutation sites were found:K^(53)→N^(53),K^(62)→R^(62),P^(183)→S^(183),G^(263)→E^(263),K^(461)→E^(461),K^(457)→R^(457),I^(475)→M^(475).Except the neutralizing antigen site G1,mutation sites were present in G2,G3 and G4.According to the results of antibody test kit,19 samples with antibody concentration≥52.5 ng/L critical value were found,with a positive rate of 31.67%.In conclusion,BEFV infection was present in this dairy farming community and there were multi-locus variants in the G gene of the prevalent strain.

关 键 词：牛流行热病毒 云南流行毒株 G基因 序列分析 

分 类 号：S823[农业科学—畜牧学]