鸭肠炎病毒强毒参考株在感染鸭体内的动态分布研究

Dynamic Distribution of Duck Enteritis Virus Standard Challenge Strain in Experimentally Infected Ducklings

关键词:  鸭肠炎病毒  实时荧光定量PCR  鸭  动态分布  

Key words:  Duck Enteritis virus  real-time fluorescent quantitative PCR  ducklings  dynamic distribution

摘要:

为了研究鸭肠炎病毒（Duck enteritis virus，DEV）强毒株在感染鸭体内的动态分布规律，根据DEV的gD基因序列设计检测引物RT-gDF和RT-gDR，建立了特异性强、敏感性高、重复性好的检测DEV的SYBR GreenⅠ实时荧光定量PCR方法。应用已建立的方法，对DEV强毒参考株感染鸭的组织脏器进行了定量检测。结果显示，接种后6 h即可在所有受检组织中检测到DEV DNA，随着病程发展，DNA拷贝数持续升高，直至接种5 d后感染鸭全部死亡。其中结肠病毒DNA拷贝数为最高，达到1013.26 copies，法氏囊、肝脏和盲肠次之，约1012 copies。本研究证实了DEV强毒对感染鸭的免疫器官、神经组织和消化系统均具有广泛的嗜性。

Abstract:

To explore the dynamic distribution of duck enteritis virus challenge standard strain (DEV CSC) in experimentally infected ducklings, the specific primers (RT-gDF and RT-gDR) were designed according to the gene DEV gD,and the SYBR GreenⅠreal-time fluorescent quantitative PCR method for virulent strain with satisfaction of specificity,sensibility and repeatability was established.Copies of DEV DNA were quantified in the collected tissues by the fluorescent quantitative PCR method.In result,DEV CSC DNA were firstly detected in all the samples at six hours after inoculation and the DNA copies subsequently continued to rise till the death of all the ducklings infected with DEV CSC virulent strain five days after inoculation. The DNA copies of DEV CSC strain in colon(1013.26)of dead ducklings was highest,and then came to bursa of Fabricius, caecumand liver, about 1012 copies. The results above comfirmedthat the virulent strain CSC had broad tissue tropism on immune organs, nerve tissue and digestive system of infected ducks.