Okada I, Matsumori A, Kawai C, Yodoi J, Tracy S
Department of Internal Medicine, Faculty of Medicine, Kyoto University, Japan.
J Mol Cell Cardiol. 1990 Sep;22(9):999-1008. doi: 10.1016/0022-2828(90)91039-a.
The presence of viral genomes in the murine heart in experimental coxsackievirus B3 myocarditis was investigated by Northern blotting analysis. Four-week-old C3H/He mice were inoculated with coxsackievirus B3. After sacrifice, the hearts were divided into 3 parts. Total RNA was extracted from one part of the heart. The other two parts were used for investigation of histopathology and of virus titer by plaque assay. The 32P-labeled cDNA probe was derived from the 5' end sequence of the coxsackievirus B3 genome. Northern blot autoradiograms of heart RNA were positive for viral RNA until day 7 and negative after day 10 and in control (uninfected) hearts. Positive autoradiograms always had the strongest signal at about 7.4 kilobase, corresponding to the size of the complete genome of the virus. The viral genomes were detected earlier than the appearance of the histopathologic changes in the murine heart. This type analysis of the viral genome in the murine myocardium may be useful in evaluating the effects of specific drugs on experimental viral myocarditis at the level of the viral genome.
通过Northern印迹分析研究了实验性柯萨奇病毒B3心肌炎小鼠心脏中病毒基因组的存在情况。将四周龄的C3H/He小鼠接种柯萨奇病毒B3。处死后，将心脏分成三部分。从心脏的一部分提取总RNA。另外两部分用于组织病理学检查和通过噬斑测定法检测病毒滴度。32P标记的cDNA探针源自柯萨奇病毒B3基因组的5'端序列。心脏RNA的Northern印迹放射自显影片在第7天之前呈病毒RNA阳性，在第10天之后以及对照（未感染）心脏中呈阴性。阳性放射自显影片在约7.4千碱基处总是具有最强的信号，这与病毒完整基因组的大小相对应。病毒基因组的检测早于小鼠心脏组织病理学变化的出现。这种对小鼠心肌中病毒基因组的分析类型可能有助于在病毒基因组水平评估特定药物对实验性病毒性心肌炎的影响。
