博尔纳病病毒接种Wistar大鼠脑组织中病毒基因组的原位检测

Detection of Borna disease virus (BDV) genome in the brain of BDV-inoculated Wistar rats by in situ hybridization

本文旨在用原位杂交法探讨博尔纳病病毒(BDV)接种Wistar大鼠脑组织中BDV基因组的分布情况。DIG RNA labeling kit标记BDV p24正链探针后,用斑点实验检测该探针的标记效率,斑点杂交法检测该探针的特异性。在其标记效率与特异性均达到实验要求后,用该探针对颅内接种BDV(H1766株)的Wistar大鼠脑组织中BDV基因组进行原位杂交检测。结果发现,接种3周后,BDV感染主要发生在皮质和海马,仅少量发生在丘脑和下丘脑;接种6周后,皮质和海马的BDV感染仍然存在,且丘脑和下丘脑的BDV感染明显增强,说明BDV在大鼠脑组织中的分布范围随着感染时间的延长而逐步扩大。本文建立的原位杂交法可用于检测BDV在Wistar大鼠脑组织内的分布与迁移情况。

The aim of the present study was to detect the Borua disease virus （BDV） genome in the brain of BDV- inoculated Wistar rats by in situ hybridization. The BDV p24 sense probe was labeled with digoxigenin （DIG） using a DIG RNA labeling kit. The labeling efficiency of the BDV p24 sense probe was conformed by spot test, and the specificity was determined by a dot hybridization assay. The BDV genome was then detected with the probe via in situ hybridization in paraffin brain sections of BDV-inoculated Wistar rats. At the third week post-BDV inoculation, the BDV genome was detected mainly in hippocampus and cortex, with minimal labeling in the thalamus and hypothalamus. Whereas at the sixth week post-BDV inoculation, the BDV genome was not only detected in hippocampus and cortex, but was also increased in thalamus and hypothalamus, suggesting that the BDV-infected region was enlarged following prolonged post-inoculation time, and that in situ hybridization with a BDV p24 sense probe can be used to observe the distribution of BDV in the inoculated Wistar rat brain.