来自巴西不同库种的狂犬溶血病毒分离株在N2a细胞中表现出不同的繁殖模式

Bruno Amorim Costa, Natalia Langenfeld Fuoco, L. Chaves, C. Adriana, I. Rodrigues, Orl, O. Ribeiro, Keila Iamamoto Nogi, K. Scheffer, I. Katz
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引用次数: 2

摘要

背景:狂犬病细胞培养感染试验是为了分离狂犬病溶血病毒,作为小鼠接种试验的替代方法。然而,街头狂犬病毒株的组织培养产生低病毒滴度。在这里,我们评估了成功分离病毒的脑组织数量,以有效地提高病毒滴度。方法:采集巴西不同储种的脑组织分离物,以不同浓度接种小鼠神经母细胞瘤细胞(N2a)。这些分离物在N2a细胞中连续传代后测定了感染病毒滴度和细胞活力。结果:接种物含狂犬溶血病毒显著,病毒滴度高,细胞无明显死亡。吸血蝙蝠维持的狂犬溶血病毒变体在N2a细胞中连续传代后,对培养系统具有显著的适应性,而狨猴分离株在N2a细胞中的繁殖模式与其他组相比明显不同。结论:基于上述结果,分离后进行病毒复制试验可用于不同病毒库的分离株,从而实现野生型病毒株的有效扩增。
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Rabies lyssavirus Isolates from Brazilian Different Reservoirs Species PresentDistinct Pattern of Propagation in N2a Cell
Background: Rabies cell culture infection test was developed for the isolation of Rabies lyssavirus and as an alternative for the mouse inoculation test. However, tissue culture for street rabies strains produces low viral titer. Here, we assessed the quantity of brain tissue for successful viral isolation toward increased virus titer in effective way. Methods: Brain tissue isolates from different reservoirs species of Brazil were harvested in different concentration and inoculated in mouse neuroblastoma cells (N2a). These isolates were measured infectious viral titer and cell viability followed by consecutive passages in N2a cells. Results: Inoculum containing were prominent Rabies lyssavirus due to higher viral titer and not significantly dead cell. After consecutive passages in N2a cells Rabies lyssavirus variant maintained by vampire bat had remarkable adaptation to the culture system, while isolates from marmoset presents distinct pattern of propagation in N2a cell when compared with other groups. Conclusion: Based on these results, the isolation followed by viral replication assay may be used in isolates from different reservoirs which enable an effective amplification of the wild type virus strains.
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