鸡传染性支气管炎病毒强毒株和弱毒疫苗株的免疫磁珠-多重RT-PCR鉴别方法的建立  

Establishment of Immunomagnetic Beads-multiplex RT-PCR Method for Identification of Virulent and Attenuated Vaccine Strains of Infectious Bronchitis Virus

作　　者：高文倩[1] 韩潇潇[1] 王红宁[1] 

机构地区：[1]四川大学生命科学学院,动物疫病防控与食品安全四川省重点实验室,“985工程”西南资源环境与灾害防治科技创新平台,生物资源与生态环境教育部重点实验室,四川成都610064

出　　处：《中国家禽》2017年第10期14-17,共4页China Poultry

基　　金：现代农业产业技术体系建设专项资金(CARS-41-K09);四川省科技支撑计划项目(2014NZ0002);国家自然科学基金项目(31372442)

摘　　要：利用鸡传染性支气管炎病毒(IBV)多克隆抗体构建的免疫磁珠富集IBV,对Gen Bank中896株IBV S1基因序列及其多变区进行分析,根据IBV强毒株和弱毒疫苗株序列差异设计引物,建立了一种基于免疫磁珠富集IBV后检测S1基因多变区的多重RT-PCR方法。结果表明:该方法能扩增出强毒株约710 bp的通用片段和约350 bp的特异片段,弱毒疫苗株能扩增出约710 bp的通用片段和约210 bp的特异片段,能鉴别出强毒株和弱毒疫苗株。免疫磁珠富集IBV后比不富集病毒用RT-PCR方法检测敏感性提高100倍。该方法为IBV检测提供了新的技术支持。Avian infectious bronchitis virus （IBV）was enriched by immunomagnetic beads constructed by polyclonal antibody against IBV. On the basis of the sequence analysis of IBV S1 gene and variable region of 896 strains in GenBank, primers were designed according to the sequence differences between virulent and attenuated vaccine IBV strains to establish a muhiplex RT-PCR method for detection of S1 gene hypervariable region after the immunomagnetic enrichment based of IBV. The results showed that the virulent strain could be amplified about 710 bp common fragment and specific fragment of about 350 bp;attenuated vaccine strain could be amplified about 710 bp common fragment and about 210 bp specific fragment. This method could distinguish virulent strain from attenuated vaccine strain. The limit of RT-PCR after immunomagnetic beads enrichment was increased by 100 times compared with that of no enrichment of IBV virus. This method provided the new technical support for IBV detection.

关 键 词：鸡传染性支气管炎病毒(IBV) 免疫磁珠 多重RT-PCR 鉴别 

分 类 号：S855.3[农业科学—临床兽医学]