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目的建立腺病毒滴度测定的TCID50方法,并比较MDCK、Vero、HEp-2及A549 4种细胞对人呼吸道腺病毒的敏感性。方法将腺病毒作连续10倍稀释,每个稀释度取100μl加入96孔细胞培养板中,随后加入消化的宿主细胞100μl,每个稀释度作10个重复,并设空白细胞培养对照。置37℃,5%CO2培养箱中。逐日观察细胞病变,并记录细胞病变孔数,直到对照细胞老化脱落为止,按Reed-Muench法计算病毒的TCID50。结果呼吸道腺病毒可在MDCK、Vero、HEp-2及A549四种细胞中增殖,并出现典型细胞病变,在观察期内A549和Vero细胞能测出TCID50值。结论四种细胞对腺病毒的敏感性依次为A549>Vero>Hep-2>MDCK。
Objective To establish a TCID50 method for the determination of adenovirus titer and to compare the sensitivity of four cell lines of MDCK, Vero, HEp-2 and A549 to human respiratory adenovirus. Methods Adenovirus was serially diluted 10-fold and 100 μl of each dilution was added to a 96-well cell culture plate followed by the addition of 100 μl of digested host cells, 10 replicates per dilution, and a blank cell culture control. Set 37 ℃, 5% CO2 incubator. The cytopathic effect was observed daily, and the number of cytopathic pores was recorded until the control cells became exfoliated. The TCID50 of the virus was calculated according to the Reed-Muench method. Results Respiratory adenovirus could proliferate in four kinds of cells, MDCK, Vero, HEp-2 and A549, and showed typical cytopathic effect. TCID50 was detected in A549 and Vero cells during the observation period. Conclusion The sensitivity of the four kinds of cells to adenovirus is A549> Vero> Hep-2> MDCK.