摘要
鸭3型腺病毒(Duck adenovirus type 3,DAdV-3)为近年来新发的以引起鸭肝脏肿大出血、肾脏大面积出血点为特征的鸭群新发疫病。为建立特异性的检测DAdV-3的PCR方法,研究通过分析鸭群中鸭腺病毒A型(DAdV-A)、鸭源禽腺病毒4型(FAdV-4)、鸭2型腺病毒(DAdV-2)和DAdV-3的Fiber 2基因特征,建立特异性检测DAdV-3的PCR方法。结果显示:优化后的PCR方法最佳退火温度为54℃,对DAdV-3扩增片段大小为548 bp;敏感性强,最低检测限为36.4 pg;特异性好,对DAdV-A、FAdV-4、鸭病毒性肠炎(DEV)、番鸭细小病毒(MDPV)、鹅细小病毒(GPV)、鸭圆环病毒(DuCV)和鹅多瘤病毒(GHPV)均无特异性扩增。表明该方法可有效用于开展新发DAdV-3的流行病学调查。
Duck adenovirus type 3(DAdV-3),which is newly identified adenovirus in ducklings,with necrosis and hemorrhage in liver and spleen.In this study,the Fiber 2 gene sequences of duck adenoviruse A(DAdV-A),fowl adenovirus 4(FAdV-4),duck adenovirus 2(DAdV-2) and DAdV-3 occurred in ducks were retrieved from GenBank in order to develop a specific PCR tool for DAdV-3 infection.Primer design software Oligo 7.0 was used to design specific primers based on the Fiber 2 gene characterization after genetic comparison.Then PCR method was established after conditional optimized.The results showed the optimal annealing temperature was 54 ℃,which only could amplify DAdV-3infection with 548 bp in length.The PCR method showed high sensitivity,with low detection limit at 36.4 pg.The PCR method was demonstrated good specificity,with no cross-amplification from common adenoviruses occurred in ducks(i.e.DAdV-A,FAdV-4) and other common duck-origin pathogens(i.e.duck enteritis virus,Muscovy duck parvovirus,goose parvovirus,duck circovirus and goose hemorrhagic polyomavirus).It′s concluded that PCR method for specific diagnosis with DAdV-3 was developed in this study,which laid foundation for further DAdV-3 epidemiological investigation.
作者
万春和
陈翠腾
施少华
程龙飞
傅光华
刘荣昌
陈红梅
傅秋玲
黄瑜
WAN Chunhe;Chen Cuiteng;SHI Shaohua;CHENG Longfei;FU Guanghua;LIU Rongchang;CHEN Hongmei;FU Qiuling;HUANG Yu(Fujian Provincial Key Laboratory for Avian Diseases Control and Prevention,Fujian Animal Diseases Control Technology Development Center,Institute of Animal Husbandry and Veterinary Medicine of Fujian Academy of Agricultural Sciences,Fuzhou,Fujian 350013)
出处
《中国家禽》
北大核心
2019年第2期20-23,共4页
China Poultry
基金
福建省农业科学院青年科技创新团队(STIT2017-3-10)
福建省属公益类项目(2018R1023-5)