柯萨奇病毒A组16型G20分离株在不同培养条件下的增殖动力学

Proliferation kinetics of Coxsackievirus A16 G20 strain under different culture conditions

目的探讨柯萨奇病毒A组16型(Coxsackievirus A16,CA16)G20分离株在不同培养条件下的增殖动力学。方法常规培养Vero和人二倍体KMB-17细胞,待细胞长至单层,接种G20株病毒,进行病毒蚀斑试验,以0.1 MOI的感染复数分别感染此两种细胞,置不同温度和pH维持液中进行培养,每2 h收样1次至第24 h,以48 h收样作为对照样,微量滴定法检测病毒滴度,并绘制增殖动力学曲线。结果 G20株病毒在Vero及KMB-17细胞中传代适应后,均可导致细胞病变。G20株病毒在此两种细胞中培养,pH 6.0、pH 6.5时,不同培养温度下,病毒基本不增殖;pH 7.5与pH 7.0的病毒增殖基本一致,增殖速度随温度呈现37℃>35℃>33℃的趋势;在33℃～37℃、pH 7.0～8.0时,病毒均有不同程度的增殖;在41℃、各pH条件下,病毒增殖均明显受到限制。结论病毒在此两种细胞中,37℃,pH 7.5培养条件下,增殖速度及滴度均较理想。

Objective To investigate the proliferation kinetics of Coxsackievirus A16（CA16） G20 strain under different culture conditions.Methods Vero and KMB-17 cells were cultured routinely till the cell monolayer was formed,on which G20 strain was inoculated for viral plaque assay.The cells were infected with G20 at a MOI of 0.1 and then cultured in medium at various temperatures and pH val ues.Samples were collected every 2 h within 24 h after infection and determined for virus titer by microtitration using that 48 h after infection as control,based on which a proliferation kinetic curve was plotted.Results Both the G20 strain adapted in Vero and KMB-17 cells caused CPE.G20 strain failed to proliferate or grew slowly at pH 6.0 and pH 6.5,of which the proliferation levels were basically identical at pH 7.5 and pH 7.0,and the proliferation rate increased with the increasing temperature（37℃ 〉 35℃ 〉 33℃）.The virus was proliferated at different degrees at 33 ～ 37℃,pH 7.0 ～ 8.0,which was restricted significantly at 41℃ with various pH values.Conclusion The proliferation rate and titer of G20 were satisfactory at 37℃,pH 7.5.