鸭瘟病毒疫苗株在免疫SPF鸭体内的生长动力学检测

Detection of the growth kinetics of duck enteritis virus vaccine strain in vaccinated SPF duck

为了研究鸭瘟病毒疫苗株在免疫SPF鸭脑、胸腺、脾、肾、肝、法氏囊中的动态分布规律,根据GenBank中鸭瘟病毒的UL6基因设计引物RTF和RTR,采用SYBR GreenⅠ实时荧光定量方法,建立了特异性强、敏感性高、重复性好的标准曲线。利用建立的标准曲线对采集组织中鸭瘟病毒DNA的拷贝数进行定量。结果显示,免疫后在脑、胸腺、肝、肾、脾、法氏囊均能检测到鸭瘟病毒DNA,且免疫后第1天至第3周,肝、脑、肾中病毒DNA的拷贝数较其他受检组织中鸭瘟病毒DNA的拷贝数高。免疫后至第9周,鸭瘟病毒DNA检出不稳定。试验结束时,除肾、法氏囊外,其他组织中病毒DNA的拷贝数均下降至1×105copies以下。表明鸭瘟病毒疫苗株对SPF鸭组织具有泛嗜性,其主要分布于SPF鸭脑、肝、肾中。

The dynamic distribution of duck enteritis virus（DEV） vaccine strain in the brain,thymus, spleen,kidney,liver and bursa of Fabricius of the SPF duck vaccinated with DEV vaccine strain were detected. The specific primers（RTF and RTR） were designed according to the gene of DEV US6 in GenBank and,the standard curve with satisfaction of specificity, sensibility and repeatability was established using SYBR Green I real-time fluorescent quantitative PCR method. Copies of DEV DNA were quantified in the collected tissues by the standard curve. In result, the DEV DNA was detected in the brain, thymus, spleen, kidney,liver and bursa of Fabricius after immunization,and the copies of DEV DNA in the liver,brain and kidney were higher than in other examined organs from the first day to the third week after immunization. Since being vaccination, the copies of DEV DNA were unstable until the ninth week. By the end of this experiment,the copies of virus DNA had fall to 1×10^5 copies in these tissues except for the kidney and bursa of Fabricius. The results showed that DEV with pantropic mainly distributed in the brain,liver and kidney.