应用SYBR GreenⅠ荧光定量PCR方法检测人工感染雏鸭体内DEV NP基因表达水平

Detection of virulent DEV NP gene in artificial infected ducklings using SYBR GreenⅠ real-time PCR

为阐明DEV的感染与致病机理,更深入地研究病原与鸭体的相互关系。将鸭肠炎病毒(DEV)强毒GZ株经腿部肌肉注射感染15日龄雏鸭后,应用建立的SYBR GreenⅠ荧光定量PCR方法对NP(病毒核衣壳蛋白)基因转录物进行定量,以检测NP基因在雏鸭体内各组织的表达水平。结果显示:NP基因在所检组织中均有表达,感染后3 h即可在肝、脾、脑、胸腺、肾和肺6种组织中检测到NP基因的表达;感染后6 h,除上述组织外还可在十二指肠和盲肠检测出;感染后10 h又可在胸肌和心肌等检测出。不同受检组织检出量有所不同,且在感染后30或48 h达到高峰,持续至96 h,之后均有所下降。

To observe the expression levels of DEV NP gene in the various organizations of 15-day-old ducklings infected intramuscularly with virulent DEV strain GZ,SYBR GreenⅠreal-time quantitative PCR was used to detect the transcripts expression levels of NP gene.The results indicated that the NP gene could be detected in six tissues including liver,spleen,brain,thymus,kidney and lung at 3 h post-infecion（PI）,and in duodenum and cecum beside the above six tissues at 6 h PI,then in pectoral muscle and cardiac muscle beside the above eight tissues at 10 h PI.The expression levels detected were various in different organizations,and reached a peak at 30 h or 48 h PI,continuing until 96 h,then all declined.These results provided a theoretical basis for clarification of pathogenic mechanism of DEV,and made the foundation for the study of the relationship between pathogen and ducks.