甲3型、乙型流感病毒在MHL混合细胞中增殖的研究

Observation of influenza A and B viruses cultured in mixed cells

〔目的〕建立MHL混合细胞瓶分离培养流感病毒的方法 ,为进行流感监测及临床快速诊断打下基础。〔方法〕采用MDCK、HEP -2、LLC混合细胞培养瓶和MDCK单一细胞培养瓶 ,同时分别接种甲 3型和乙型流感病毒标准毒株 ,并设立阴性细胞对照组 ,观察甲 3型和乙型流感病毒毒株在混合细胞瓶和MDCCK细胞瓶中的细胞病变效应 (CPE)、血凝滴度和单克隆抗体荧光染色情况。〔结果〕病毒血凝滴度分别均为 1:160 ,经 1:5稀释的甲 3型和乙型流感病毒接种MHL混合细胞瓶和MDCK细胞瓶经 3 7℃、48h培养后 ,感染甲 3型病毒细胞出现细胞拉网、团聚病变现象 ,感染乙型流感病毒细胞则出现明显细胞固缩、脱落现象 ,MDCK单一细胞培养瓶CPE强于混合细胞培养瓶 :3 7℃、72h培养后 ,检测血凝滴度 ,感染甲 3型流感病毒的MDCK细胞和混合细胞均为l:40。感染乙型流感病毒的MDCK细胞为 1:12 0 ,MHL混合细胞为 1:2 0 ;单克隆荧光抗体检测 ,感染甲 3型和乙型流感病毒的MDCK细胞和混合细胞均呈阳性。〔结论〕应用MDCK、HEP-2 ,LLC混合细胞瓶分离、培养 ,不仅可对甲型流感爆发进行流行病学监测 ,同时还可分离鉴定其它呼吸道病毒 ,利于临床诊断、治疗和预防。

We inoculate influenza A (H3) and B virus in a shell-vial of mixed cells of MDCK, Hep-2 and LLC-MK2 cells or shell vials of MDCK cells alone to observe for the presence of cytopathic effect (CPE), haemagglutination and monoclonal antibody-based immunofluoreseene assay Results demonstrated that after incubated at 37℃ for 24h, the mixed cell culture or MDCK cells alone which were inoculated with influenza A virus, cytopathic effect of viral infection were observed The CPE observed in the MDCK cells alone vial was stronger than that of the mixed cell culture After incubated at 37℃ for 72h, hemagglutination assay showed that tbr influenza A (H3) virus, the titres were both 1:40 in mixed cells vial and MDCK alone vial while influenza B virus inoculated in MDCK cells vial is l:120 and inoculated in mixed cells vial is 1:20 Influenza A (H3) and B virus were detected in mixed cells vial or in MDCK cells vial by McAb-based immunofiuorescence assay