运用实时荧光定量PCR法检测感染鸡种蛋中的REV  

Detection of REV in Serum and Eggs of Infected Chickens by Real-time Fluorescent Quantitative PCR

作　　者：黄小洁[1] 杨承槐[1] 刘丹[1] 侯力丹[1] 李慧姣[1] 李俊平[1] 陈晓春[1] 

机构地区：[1]中国兽医药品监察所,北京100081

出　　处：《中国兽药杂志》2017年第11期15-19,共5页Chinese Journal of Veterinary Drug

基　　金：国家重点研发计划项目"禽病检测标准物质与计量技术方法研究"课题(2016YFD0500809)

摘　　要：为研究通过种蛋检测的方法监测SPF鸡群REV感染情况,运用已建立的荧光定量PCR方法检测攻毒母鸡种蛋中REV,并与血清REV水平进行比较。人工感染12月龄母鸡,从种蛋中提取蛋清,接种正常鸡胚成纤维细胞后传代一次,之后石蜡密封孵化至9日龄制成鸡胚成纤维细胞(CEF),运用荧光定量PCR方法检测蛋清,并和血清的检测结果进行比较,结果显示:83份蛋清,3份阳性样品分别收集自攻毒后的第8、11、14天;46份CEF,2份阳性样品分别收集自攻毒后的第11天和第14天,阳性的蛋清和CEF呈对应关系,说明两种方法的检测结果一致。攻毒后的6~15 d是病毒血症持续期,阳性鸡蛋均采集自这段时间,表明通过种蛋REV检测能反映SPF鸡的急性感染状态。本研究为通过种蛋抗原检测从而监测SPF鸡REV感染奠定基础。The aim of this study was to study the egg detection indicating the REV infection status in SPF chickens. The REV of eggs in infected hens was detected by real-time PCR method already established, and compared with the serum. 12 month-old hens inoculated with REV.The egg white was extracted from the eggs and was inoculated into chicken embryo fibroblast（CEF） and passaged once, then these eggs were sealed with paraffin wax and hached, CEF was made from 9 days embryos. These egg white and CEF were detected by the real-time PCR then compared with the serum. The result showed from 83 samples, which collecting from the eighth and that REV in three egg white samples could be detected the eleventh and the fourteenth day after infection. Two CEF samples could be detected from 46 samples, which collecting from the eleventh and the fourteenth day after infection. The positive egg white and CEF was correspondence. The result indicated the two methods were consistent. The viremia continued from 6 to 18 days after attack and the positive eggs were collected from this time. The result indicated that the egg detection could be used to monitor the acute infection status in SPF chicken. The research provided a foundation for monitoring REV infection in SPF chickens by egg antigen test.

关 键 词：网状内皮组织增生症病毒 荧光定量PCR 血清 蛋清 鸡胚成纤维细胞 

分 类 号：S852.65[农业科学—基础兽医学]