Dangler C A, de la Concha-Bermejillo A, Stott J L, Osburn B I
Department of Pathology, School of Veterinary Medicine, University of California, Davis 95616.
J Vet Diagn Invest. 1990 Oct;2(4):303-7. doi: 10.1177/104063879000200409.
In situ nucleic acid hybridization was tested for the ability to detect bluetongue virus (BTV) nucleic acids in blood mononuclear cells. A standard protocol was devised and applied to the demonstration of BTV genetic sequences in cultured bovine mononuclear cells that had been infected in vitro. In situ hybridization using biotinylated single-stranded RNA probes, in the presence of 50% formamide at 50 C, demonstrated an intense, positive signal in 0.001-0.01% of the BTV-infected cultured mononuclear cells. The protocol was applied to isolated mononuclear cells from an experimentally infected heifer. No infected cells were observed by this method, although the blood specimens were obtained during peak viremia.
对原位核酸杂交检测血液单核细胞中蓝舌病毒（BTV）核酸的能力进行了测试。设计了一种标准方案并将其应用于在体外感染的培养牛单核细胞中检测BTV基因序列。使用生物素化单链RNA探针，在50℃、50%甲酰胺存在的条件下进行原位杂交，结果显示在0.001%-0.01%的BTV感染培养单核细胞中出现强烈的阳性信号。该方案应用于从一头实验感染的小母牛分离的单核细胞。尽管在病毒血症高峰期采集了血液样本，但通过该方法未观察到感染细胞。
