Department of Neurovirology, WHO Collaborating Centre for Reference and Research on Rabies, National Institute of Mental Health and Neurosciences (NIMHANS), Hosur Road, Bangalore, India.
Int J Infect Dis. 2010 Dec;14(12):e1067-71. doi: 10.1016/j.ijid.2010.07.004. Epub 2010 Oct 13.
A confirmatory rabies diagnosis can be achieved by rapid virus isolation in cell culture using brain tissue from the suspect animal. Several cell lines have been used for this purpose and the murine neuroblastoma cell line Neuro-2a has been found to be the most sensitive. The human embryonic kidney cell line HEK-293 is known to express several neuronal proteins and is believed to be of neuronal origin. We hypothesized that this cell line could be susceptible to rabies virus, which is highly neurotropic.
First we tested the sensitivity of HEK-293 cells to the laboratory strain, challenge virus standard (CVS). We then tested 120 brain samples from different animals and humans suspected to have died of rabies by fluorescent antibody test (FAT). Both FAT-positive and FAT-negative brains were tested for virus isolation using Neuro-2a, BHK-21, and HEK-293 cell lines and also by mouse inoculation.
There was 100% correlation between FAT, virus isolation in Neuro-2a and HEK-293 cells, and mouse inoculation. However, the rate of virus isolation in the BHK-21 cell line was only 28% when compared to the other cell lines. The sensitivity of HEK-293 to CVS strain of virus was similar to that of Neuro-2a.
We conclude that the HEK-293 cell line is as sensitive as the Neuro-2a cell line for the rapid isolation of rabies virus and may serve as an alternative cell line for rabies diagnosis and future research.
通过使用来自可疑动物的脑组织在细胞培养物中快速分离病毒,可以确诊狂犬病。为此目的已经使用了几种细胞系,其中鼠神经母细胞瘤细胞系 Neuro-2a 被发现是最敏感的。人胚肾细胞系 HEK-293 已知表达几种神经元蛋白,据信其来源于神经元。我们假设该细胞系可能易受高度嗜神经的狂犬病病毒感染。
首先,我们测试了 HEK-293 细胞对实验室株、挑战病毒标准株(CVS)的敏感性。然后,我们用荧光抗体试验(FAT)测试了来自不同动物和疑似死于狂犬病的人类的 120 个脑组织样本。使用 Neuro-2a、BHK-21 和 HEK-293 细胞系以及小鼠接种,对 FAT 阳性和 FAT 阴性的脑组织进行病毒分离测试。
FAT、Neuro-2a 和 HEK-293 细胞系以及小鼠接种的病毒分离之间存在 100%的相关性。然而,与其他细胞系相比,BHK-21 细胞系的病毒分离率仅为 28%。HEK-293 对 CVS 株病毒的敏感性与 Neuro-2a 相似。
我们得出结论,HEK-293 细胞系与 Neuro-2a 细胞系一样,可用于快速分离狂犬病病毒,并且可以作为狂犬病诊断和未来研究的替代细胞系。