摘要
旨在建立一种快速、准确检测H4亚型禽流感病毒(AIV)的方法,针对H4亚型AIV HA基因保守序列,设计并筛选出2对特异性引物,通过优化反应条件,建立了H4亚型AIV套式RT-PCR检测方法。特异性试验结果显示,该法可特异性检测出H4亚型AIV,对其他常见禽病病原体无交叉;敏感性试验结果显示,该法对H4亚型AIV检测下限为360fg/μL;用该法对106份临床样品进行检测,临床样品检测结果与病毒分离结果一致。本研究建立的套式RT-PCR方法为H4亚型AIV的早期诊断及有效防控提供了快速、特异和敏感的检测方法。
In order to develop a simple and quick method to detect H 4 subtype of avian influenza virus （AIV） ,two pairs of primers were designed based on the HA gene sequences of AIV H4 subtype .A nested RT‐polymerase chain reaction （nRT‐PCR） method was established for detection of H4 subtype of AIV .To further examine reliability ,specificity and sensitivity of the assay were evaluated ,and 106 clinical samples were tested by nested RT‐PCR .The results show that this nested RT‐PCR was specific to the H4 subtype of AIV ,without amplification of genomic materials other avian pathogens ;The detection limit of nested RT‐PCR was 36 .0 fg/μL ;The results of nRT‐PCR for detection of clinical samples were coincident with that of the viral isolation completely .Our results demonstrated that the optimized nested RT‐PCR assay was quick ,specific and sensitive for detection of H4 subtype of AIV .
出处
《动物医学进展》
北大核心
2015年第2期11-15,共5页
Progress In Veterinary Medicine
基金
广西特聘专家专项(2011B020)
广西科技重大专项项目(14121003-4-2)
广西畜禽疫苗新技术重点实验室项目(13-051-27-A-2)
