牧羊犬外周血和脑组织博尔纳病病毒检测结果的比较

Borna disease virus detection in peripheral blood and brain tissues of shepherd dogs

目的检测同一牧羊犬外周血和脑组织博尔纳病病毒(BDV)p24片段,比较两者阳性率的差异。方法采用改进的荧光定量套式RT-PCR,对新疆伊犁地区圈养的100只牧羊犬外周血单核细胞(PBMC)和脑组织(BT)同时进行BDV p24基因片段的检测,并对阳性标本采用GFP-p24、pMD-19扩增后电泳验证,排除质粒污染,并克隆测序,用Fisher精确检验和χ2检验分析两者阳性率的差异。结果牧羊犬外周血单核细胞和脑组织BDV p24检测的阳性率分别为5%和9%;PBMC组和BT组BDV p24检测阳性率差异无统计学意义(P>0.05)。结论BDV在脑内持续感染,但以RT-PCR对外周血单核细胞BDV p24片段的检测有可能替代脑组织BDV检测,作为大规模流行病学调查的手段。

To detect and analyze the differences of Borna disease virus （BDV） p24 gene segment in peripheral blood mononuclear cell（PBMC）and brain tissue（BT） of animals, the BDV p24 gene segments were detected by the modified fluorescent quantitative real-time PCR in peripheral blood and in brain tissues of 100 shepherd dogs in Ili area, Xinjiang. After amplifying and excluding plasmid contamination, the positive samples were validated by electrophoresis and then sequenced. Fisher’s test and χ2 test demonstrated that the positive rates in peripheral blood and brain tissue were 5% and 9%, respectively. There was no statistically significant difference between them in p24 gene（P0.05）. It was apparent that modified fluorescent quantitative real-time PCR could be used to detect the BDV p24 gene segments in peripheral blood, which might reflect the infection in brain tissue. It also suggested that the detection of peripheral blood for BDV infection would take place of brain tissue detection in large-scale epidemiological survey.