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目的了解H7N9流感病毒在不同细胞中增殖的感染动力学差异。方法将H7N9流感病毒分别接种于鸡成纤维细胞(DF-1)、狗肾细胞(MDCK)、人支气管上皮细胞(BEAS-2B)、人肺腺癌细胞(A549),观察病毒在各细胞内增殖后细胞病变效应(CPE)、血凝实验滴度及50%组织培养细胞感染剂量。结果 H7N9流感病毒在A549细胞中培养36 h可达到血凝效价最高,同时可见明显CPE,在DF-1及MDCK细胞中培养72 h后血凝效价达到最高;在BEAS-2B细胞中增殖不明显,培养液对豚鼠血无明显凝集,且培养超过96 h尚无明显CPE。DF-1细胞、MDCK细胞、A549细胞对病毒的TCID50实验结果分别为10~(-4.34)、10~(-5.13)、10~(-3.84)。结论本实验使用的H7N9流感病毒不能有效感染BEAS-2B细胞并在其中增殖,但是可以在DF-1、MDCK及A549细胞中有效增殖。增殖能力以MDCK细胞最强,DF-1细胞略弱,A549细胞次之。A549细胞对病毒的吸附能力较强,可快速达到增殖的高峰。
Objective To understand the kinetics of infection in H7N9 influenza virus that proliferate in different cells. Methods Human H7N9 influenza virus was inoculated into DF-1, MDCK, BEAS-2B and A549 cells respectively. Proliferative cytopathic effect (CPE), titer of hemagglutination test and infection dose of 50% tissue culture cells. Results The H7N9 influenza virus could reach the highest titer of hemagglutination and obvious CPE when cultured in A549 cells for 36 h. The titer of hemagglutination reached the highest in 72 h after cultured in DF-1 and MDCK cells, and proliferated in BEAS-2B cells Obviously, the culture medium had no obvious agglutination on the guinea pig blood, and there was no obvious CPE when cultured for more than 96 h. The results of TCID50 of DF-1 cells, MDCK cells and A549 cells were 10 ~ (-4.34), 10 ~ (-5.13), 10 ~ (-3.84), respectively. Conclusion The H7N9 influenza virus used in this experiment failed to effectively infect and proliferate BEAS-2B cells, but effectively proliferated in DF-1, MDCK and A549 cells. The proliferation of MDCK cells strongest, DF-1 cells slightly weaker, followed by A549 cells. A549 cells strongly adsorb the virus and rapidly reach the peak of proliferation.