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目的对不同温度培养的3株EV71中国流行株的增殖动力学进行分析。方法分别在32、35、37和40℃条件下对FY23、K9和S1株EV71进行传代培养、2～24h培养及噬斑培养,采用微量滴定法测定病毒的感染性滴度;传代培养的子代病毒通过脑内注射方式感染新生ICR乳鼠,观察病毒的致病性。结果传代培养时,在32和40℃培养的3株病毒致细胞完全病变时间较35和37℃延长,且子代病毒滴度下降,35℃培养的病毒滴度较37℃升高;3株病毒培养24h后,37℃培养的病毒增殖效率最高,35℃次之,40℃最低;4个温度培养的病毒噬斑形态无明显变化;新生乳鼠脑内注射致死率差异无统计学意义(P>0.05)。结论培养温度的变化可影响EV71的致细胞病变速度及子代病毒的增殖,但对病毒噬斑的形态及新生乳鼠的致病能力无明显影响。
Objective To analyze the proliferation kinetics of three Chinese strains of EV71 cultured at different temperatures. Methods The EV71 cells of FY23, K9 and S1 strains were subcultured at 32, 35, 37 and 40 ℃ for 2 ~ 24h culture and plaque culture respectively. The infectivity titer of the virus was determined by microtiter method. Generation of virus infection by intracerebral injection of neonatal ICR suckling mice to observe the pathogenicity of the virus. Results In the subculture, the complete pathogenicity of three strains of cells cultured at 32 and 40 ℃ was longer than that of 35 and 37 ℃, and the titer of progeny virus was decreased. The titer of virus at 35 ℃ was higher than that at 37 ℃. After culturing for 24 hours, the proliferation efficiency of virus at 37 ℃ was the highest, followed by 35 ℃ and lowest at 40 ℃. There was no significant change in virus plaque morphology at 4 temperature. There was no significant difference in injection rate of neonatal neonatal rats P> 0.05). Conclusion The change of culturing temperature can affect the rate of cytopathic effect of EV71 and the proliferation of progeny virus, but has no obvious effect on the morphology of viral plaque and the virulence of neonatal rats.