2株云南源BVDV-1b亚型毒株的分离和鉴定  

Isolation and Identification of Two BVDV-1b Subtype Strains from Yunnan Province

作　　者：崔垚鑫 李聃 谢佳芮 朱沛 徐心明 岩锐 陈其云 李文贵 姚俊[2] CUI Yaoxin;LI Dan;XIE Jiarui;ZHU Pei;XU Xinming;YAN Rui;CHEN Qiyun;LI Wengui;YAO Jun(College of Veterinary Medicine,Yunnan Agricultural University,Kunming 650201,China;Yunnan Provincial Key Laboratory of Tropical and Subtropical Animal Viral Diseases,Yunnan Academy of Animal Science,Kunming 650224,China;Dali Town Agricultural Comprehensive Service Center,Dali City,Yunnan Province,Dali 671003,China;Lufeng Center for Animal Disease Control and Prevention,Chuxiong Prefecture,Yunnan Province,Lufeng 651200,China;Center for Animal Disease Control and Prevention of Menglian County,Pu'er City,Yunnan Province,Pu'er 665000,China;College of Animal Science and Technology Yuxi Agricultural Vocational and Technical College,Yuxi 651100,China)

机构地区：[1]云南农业大学动物医学院,云南昆明650201 [2]云南省畜牧兽医科学院云南省热带亚热带动物病毒病重点实验室,云南昆明650224 [3]云南省大理市大理镇农业综合服务中心,云南大理671003 [4]云南省楚雄州禄丰市动物疫病预防控制中心,云南禄丰651200 [5]云南省普洱市孟连县动物疫病预防控制中心,云南普洱665000 [6]玉溪农业职业技术学院动物科技学院,云南玉溪651100

出　　处：《中国兽医杂志》2025年第2期40-46,共7页Chinese Journal of Veterinary Medicine

基　　金：云南省科技厅乡村振兴科技专项-科技特派团(202304BI090003);云南省科技厅重点研发计划(202303AK140031);云南益民养殖示范基地有限公司姚俊专家基层科研工作站;澜沧重牧农业科技有限公司姚俊专家基层科研工作站。

摘　　要：为探明云南省牛病毒性腹泻黏膜病(BVD-MD)的发生、流行态势及其流行毒株的生物学特征,本试验于2022年12月—2023年11月从云南省10个州市规模化养牛场随机采集667份牛粪便样品,进行牛病毒性腹泻病毒(BVDV)逆转录-聚合酶链反应(RT-PCR)检测,针对阳性样品选用牛肾细胞(MDBK)进行毒株的分离和鉴定,结合间接免疫荧光试验(IFA)观察病毒增殖情况,并使用Reed-Muench法计算病毒滴度;阳性样品经全基因组测序后,分别对全基因组和5′非翻译区(5′-UTR)序列进行系统发育分析。结果显示,经RT-PCR和MDBK细胞分离鉴定获得2株BVDV,分别命名为BVDV YN.1(GenBank登录号:PP488512.1)和BVDV YN.2(GenBank登录号:PP770551.1),病毒滴度分别为105.81和104.93 TCID50/mL。系统发育分析结果显示,2株BVDV分离株与美国分离的BVDV-1b亚型参考毒株HP-KY-RK13(GenBank登录号:KT355592.1)的核苷酸相似度分别为99.6%和99.5%,与日本分离的BVDV-1b亚型参考毒株END(GenBank登录号:JX419397.1)的核苷酸相似度均为99.7%,且处于同一进化分支(BVDV-1b亚型)。本试验可为云南省BVDV流行病学调查及对其防控提供理论参考依据。To investigate the occurrence,epidemiological trends,and biological characteristics of circulating strains of bovine viral diarrhea mucosal disease(BVD-MD)in Yunnan Province,this study randomly collected 667 bovine fecal samples from largescale cattle farms across 10 prefectures in Yunnan between December 2022 and November 2023.The samples were tested for bovine viral diarrhea virus(BVDV)using reverse transcription-polymerase chain reaction(RT-PCR).Positive samples were used to isolate and identify strains on Madin-Darby bovine kidney(MDBK)cells,with virus propagation monitored through indirect immunofluorescence assay(IFA).Virus titers were determined using the Reed-Muench method.Whole-genome sequencing was performed on positive isolates,and phylogenetic analyses were conducted based on full-genome and 5′untranslated region(5′-UTR)sequences.The results showed that two BVDV strains were successfully isolated and identified,named BVDV YN.1(GenBank accession No.:PP488512.1)and BVDV YN.2(GenBank accession No.:PP770551.1),with virus titers of 105.81 and 104.93 TCID50/mL,respectively.Phylogenetic analysis revealed that these two isolates shared 99.6%and 99.5%nucleotide similarity with the BVDV-1b subtype reference strain HP-KY-RK13(GenBank accession No.:KT355592.1)from the United States and 99.7%similarity with the BVDV-1b reference strain END(GenBank accession No.:JX419397.1)from Japan,clustering within the same evolutionary branch of the BVDV-1b subtype.This study provides theoretical insights for the epidemiological investigation and control of BVDV in Yunnan Province.

关 键 词：牛病毒性腹泻病毒(BVDV) BVDV-1b亚型 云南 分离和鉴定 

分 类 号：S885.3[农业科学—特种经济动物饲养]