鸡六种病毒多重PCR检测方法的建立和应用  

Establishment and Application of a Multiplex PCR Assay for Simultaneous Detection of six Chicken-related Virus

作　　者：吴静[1,2] 赵润鹏[1] 徐雪维 王婉[1] 杨小康[1] 胡东[1,2] 

机构地区：[1]安徽理工大学医学院免疫与检验教研室,安徽淮南232001 [2]安徽理工大学免疫与感染研究所,安徽淮南232001

出　　处：《安徽理工大学学报（自然科学版）》2016年第3期8-12,共5页Journal of Anhui University of Science and Technology:Natural Science

基　　金：国家自然科学基金资助项目(81571528;81202294;81302524);安徽省高校自然科学产学研基金资助项目(KJ2013A105);安徽省自然科学基金资助项目(1208085QH162);国家级大学生创新创业训练计划基金资助项目(201210361112;201210361121);安徽理工大学青年科技拔尖人才基金资助项目

摘　　要：禽流感(Avian influenza,AI)、鸡新城疫(Newcastle disease,ND)、鸡传染性支气管炎(Infectious bronchitis,IB)、鸡传染性喉气管炎(Infectious laryngotracheitis,ILT)、鸡传染性法氏囊病(Infectious bursal disease,IBD)和鸡减蛋综合症(Egg drop syndrome 76,EDS-76)是6种常见的禽类病毒性传染病,给养鸡业造成重大经济损失。依据NCBI核酸数据库中禽流感病毒(Aiv)、新城疫病毒(Ndv)、鸡传染性支气管炎病毒(Ibv)、传染性法氏囊病(Ibdv)、鸡传染性喉气管炎病毒(Iltv)及减蛋综合征病毒(Edsv)基因序列,设计了6对特异性引物,建立并优化针对鸡Aiv、Ndv、Ibv、Ibdv、Iltv和Edsv的多重PCR鉴别诊断方法,同时进行现场检测应用。结果显示该多重PCR体系检测限为100fg,与其他常见病原体无交叉反应。研究表明鸡六种病毒多重PCR方法具有较高敏感性与特异性,在禽类常见病毒现场诊断方面具有广阔的应用前景。Avian influenza virus （Aiv） , New castle disease virus （Ndv） , Infectious bronchitis virus （Ibv） , In- fectious laryngo tracheitis virus （Iltv）, Infectious bursal disease virus （Ibdv） and Egg drop syndrome virus （Edsv） combine to be the six common chicken -related viruses, which pose great threat to poultry industry. Ac- cording to the gene sequences of the six common viruses in Nucleotide GenBank, six pairs of specific primers were designed and a multiplex PCR for simultaneous detection of the six viruses was established and optimized. At the same time, some clinical samples were detected. The result shows that this method is performed with a de- tection limit of 100 fg which had no cross reaction to other pathogens. This study indicates that this method has a good sensitivity and specificity, and it may have an expanded application to the diagnosis of viral diseases in poultry.

关 键 词：多重PCR 禽流感病毒 新城疫病毒 鸡传染性支气管炎病毒 鸡传染性喉气管炎病毒 

分 类 号：S831.7[农业科学—畜牧学]