区别clade 2.3.2.1和clade 2.3.4.4 H5亚型禽流感病毒实时荧光定量PCR方法的建立与应用  

Establishment and application of real-time quantitative PCR for differentiation of clade 2.3.2.1 and clade 2.3.4.4 H5 avian influenza

作　　者：陈珍[1,2] 朱春华 陈翠腾[1,2] 刘斌琼 蔡国漳[1,2] 施少华 万春和[1,2] 黄瑜 Chen Zhen;Zhu Chunhua;Chen Cuiteng;Liu Bingqiong;Cai Guozhang;Shi Shaohua;Wan Chunhe;Huang Yu(ABSL-3 of Institute of Animal Husbandry and Veterinary,Fujian Academy of Agricultural Sciences,Fuzhou350013;Fujian Provincial Key Laboratory for Avian Diseases Control and Prevention,Fuzhou350013;Institute of Biotechnology,Fujian Academy of Agricultural Sciences,Fuzhou350003)

机构地区：[1]福建省农业科学院畜牧兽医研究所ABSL-3实验室,福州350013 [2]福建省禽病防治重点实验室,福州350013 [3]福建省农业科学院生物技术研究所,福州350003

出　　处：《福建畜牧兽医》2024年第6期23-26,共4页Fujian Journal of Animal Husbandry and Veterinary medicine

基　　金：福建省属公益类科研院所基本科研专项(2021R10260012);福建省自然科学基金面上项目(2022J01466);财政部和农业农村部:国家现代农业产业技术体系项目(CARS-42);福建省财政专项(FJFS-2023)资助。

摘　　要：禽流感(Avian influneza,AI)是由正粘病毒科的禽流感病毒(AIV)引起的,疫苗接种策略是控制高致病性禽流感的有效手段,而疫苗接种策略成功的关键在于有与流行毒株相匹配的疫苗株。本试验根据clade 2.3.2.1和clade 2.3.4.4 H5 AIV血凝素(HA)基因所存在的特征性核苷酸序列差异来设计1对引物,利用实时荧光定量PCR反应后生成熔解曲线的Tm值与其GC含量正相关的特点,通过观察实时荧光定量PCR扩增反应后熔解曲线Tm值差异,即可精确对clade 2.3.2.1和clade 2.3.4.4 H5AIV感染情况进行准确鉴别诊断,为H5 AIV疫苗使用提供科学数据。Avian influenza(AI)is caused by avian influenza virus(AIV),a member of the Orthomyxovirus family,Vaccination strategy is an effective mean of controlling highly pathogenic avian influenza,and the key to a successful vaccination strategy is to have a vaccine strain that matches the prevailing strain.In this study,a pair of real-time PCR primers was designed according to the characteristic nucleotide sequence difference between clade 2.3.2.1 and clade 2.3.4.4 H5 AIV hemagglutinin(HA)gene,based on the positive correlation between the Tm value of the melt curve generated by real-time quantitative fluorescent PCR reaction and its GC content,differential diagnosis of clade 2.3.2.1 and clade 2.3.4.4 H5 AIV infection can be made by observing the difference of the Tm value of the melt curve of real-time quantitative PCR reaction,the result would be accessible way to provide scientific data for the use of H5 AIV vaccine.

关 键 词：禽流感病毒 H5亚型 Clade2.3.2.1 Clade2.3.4.4 鉴别诊断 

分 类 号：S852.65[农业科学—基础兽医学]