犬肾细胞系、人喉表皮癌细胞、非洲绿猴肾细胞混合培养检测常见呼吸道病毒和肠道病毒  

Mixed culture of Madin-Darby canine kidney cells, human epidermoid cancer cells and African green monkey kidney cells for detection of common respiratory viruses and enteroviruses

作　　者：高颖阳[1] 姜晨彦[2] 蒋露芳[1] 汪千力[1] 居丽雯[1] 

机构地区：[1]复旦大学公共卫生学院流行病学教研室公共卫生安全教育部重点实验室,上海200032 [2]上海市疾病预防控制中心

出　　处：《中华传染病杂志》2011年第6期321-324,共4页Chinese Journal of Infectious Diseases

基　　金：“十一五”国家重大专项（2009ZX10004-104）;国家高技术研究发展计划（863计划）项目（2007AA02Z407）

摘　　要：目的建立犬肾细胞系（MDCK）、人喉表皮癌细胞（Hep-2）、非洲绿猴肾细胞（Vero）混合培养以分离常见呼吸道病毒和肠道病毒，为临床检测主要呼吸道、肠道病毒提供新方法。方法采用MDCK、Hep-2和Vero制备混合瓶，接种甲型流行性感冒（流感）病毒、乙型流感病毒、腺病毒和肠道病毒71型流行株污染的咽拭子，观察细胞病变效应，以及采用单克隆荧光抗体、RT—PCR和PCR鉴定。结果对应不同病毒，敏感细胞系出现明显的细胞病变效应，荧光抗体检测可见特异的绿色荧光颗粒，PCR产物电泳结果可见不同的目的片段。结论混合细胞瓶一次可以分离培养多种常见呼吸道病毒和肠道病毒，通过后续的特异性检测手段即可检测，比直接检测标本抗原、核酸的检出率高。该方法有利于推动临床病毒性疾病的诊断。Objective To establish a clinical test assay for detecting common respiratory viruses and enteroviruses （EV） by using mixed cultured Madin-Darby canine kidney cells （MDCK）, human epidermoid cancer cells （Hep-2） and African green monkey kidney cells （Vero） to isolate common respiratory viruses and enteroviruses. Methods Throat swabs with influenza A and B viruses, adenovirus and EV71 were incubated with mixed cultured MDCK, Hep-2 and Veto in a single vial to observe the presence of cytopathie effects. Polymerase chain reaction （PCR）, reverse transcription （RT）-PCR and monoclonal antibody-based immunofluorescene assay were also used for confirmatory test. Results The sensitive cell lines developed obvious cytopathic effects to the corresponding viruses, which were confirmed by the specific green particles observed by immunofluorescence assay and specific target PCR segments. Conclusions The shell-vial of mixed cells can simultaneously isolate different common respiratory viruses and EV. The isolated pathogens can be further confirmed by antigen test and PCR. This assay may improve the diagnosis of clinical viral diseases.

关 键 词：细胞 培养的 呼吸系病毒 肠道病毒属 肾 细胞系 喉 

分 类 号：R[医药卫生]