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目的探讨国内首次分离的南定病毒致白纹伊蚊细胞病变情况。方法用长成单层的白纹伊蚊细胞感染本课题组在国内首次发现的南定病毒,观察细胞病变情况和进行分子生物学检测。结果南定病毒接种白纹伊蚊细胞后,7d开始出现病变,细胞固缩,单层细胞稀疏,细胞之间空隙增大,直到细胞数量减少,细胞间空隙增大,单层消失,空白对照生长正常,TCID50为1.41×10-6PFU/ml。应用TaqMan-MGB Probe Real Time PCR技术,检测细胞病变孔的培养液,荧光PCR结果均出现“S”扩增曲线,显示均存在NDiV核酸。用TaqMan-MGB Probe Real Time PCR制作相对定量标准曲线,6个稀释度的Ct值标准曲线R2≈1.0,呈线性关系。结论南定病毒能够引起白纹伊蚊细胞出现明显细胞病变效应。
Objective To investigate the cytopathic effect of Nanding virus on Aedes albopictus isolated for the first time in China. Methods The monolayer Aedes albopictus cells were infected with the Nanking virus, which was first discovered in our country, to observe the cytopathic effect and to perform molecular biology tests. Results After S. aureus was inoculated with Aedes albopictus, lesions began to appear on the 7th day. Cell shrinkage was observed. The monolayer cells became sparse and the interstitial spaces increased. The number of cells decreased, the intercellular space increased, the monolayer disappeared, and the blank control Growth is normal, TCID50 is 1.41 × 10-6PFU / ml. The TaqMan-MGB Probe Real Time PCR was used to detect the cells in the cytopathic cells. Fluorescence PCR showed “S” amplification curves, which showed NDiV nucleic acids were all present. Relative quantitative standard curve was generated by TaqMan-MGB Probe Real Time PCR. The calibration curve of six dilutions Ct was R2 ≈1.0, showing a linear relationship. Conclusion Nanding virus can cause significant cytopathic effects of Aedes albopictus cells.