Ⅰ群禽腺病毒套式PCR检测方法的建立与应用  

DEVELOPMENT AND APPLICATION OF NESTED PCR FOR DETECTION OF FOWL ADENOVIRUS GROUP Ⅰ

作　　者：朱小甫[1] 吴旭锦[1] 高睿[2] 尚红梅 ZHU Xiao-fu;WU Xu-jin;GAO Rui;SHANG Hong-mei(Animal Epidemic Disease Diagnostic Laboratory of Molecular Biology, Institute of Animal Husbandry and Veterinary Medicine, Xianyang Vocational Technical College, Xianyang 712000, China;Yangling Vocational Technical College, Yangling 712000, China;Xianyang City Animal Disease Prevention and Control Center, Xianyang 712000, China)

机构地区：[1]咸阳职业技术学院畜牧兽医研究所动物疫病分子生物学诊断实验室,咸阳712000 [2]杨凌职业技术学院,杨凌712100 [3]咸阳市动物疫病预防控制中心,咸阳712000

出　　处：《中国动物传染病学报》2019年第3期74-77,共4页Chinese Journal of Animal Infectious Diseases

基　　金：陕西省科技计划项目(2017NY-097);陕西省教育厅科学研究项目(17JK1171);咸阳市科技计划项目(2016k02-54);咸阳职业技术学院2017年科学技术研究重大项目(2017KYA01)

摘　　要：本研究根据GenBank发表的Ⅰ群禽腺病毒(Fowl adenovirus,FAdV)基因序列,设计了2对通用检测引物,通过反应体系和条件优化,建立了Ⅰ群禽腺病毒套式PCR检测方法。通过灵敏性检测、特异性试验以及对临床病料中FAdV检测验证实用性与可靠性。建立的套式PCR检测方法灵敏度为3.12×10-6 ng/μL;特异性试验证实该方法仅能从FAdV参考阳性株扩增出750 bp的预期目的条带,其他5种常见禽病毒均为阴性。结果表明,本研究成功建立了一种灵敏度高、特异性好的检测Ⅰ群禽腺病毒通用套式PCR方法,为临床FAdV感染的快速准确诊断提供了技术手段。According to the published gene sequences of fowl adenovirus(FAdV), two pairs of primers were designed and synthesized for development of a universal nested PCR method. The PCR method was optimized for its reaction system and conditions, sensitivity and specificity. The results showed that the detection limit of the method was 3.12×10-6 ng/μL of FAdV DNA. The target band of 750 bp was amplified from the reference fowl adenovirus but not other five avian viruses. The development of the nested PCR provided a useful tool for detection of FAdV.

关 键 词：群禽腺病毒 Hexon基因 套式PCR 检测 

分 类 号：S852.659.1[农业科学—基础兽医学]