混合细胞瓶对临床标本中流行性感冒病毒和肠道病毒的分离检测  

Mixed cells in shell vials for detection of influenza viruses and enteroviruses from clinical specimens

作　　者：汪千力[1] 熊海燕[1] 申惠国[2] 居丽雯[1] 蒋露芳[1] 蔡韵[1] 姜庆五[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]上海市闵行区疾病预防控制中心

出　　处：《中华传染病杂志》2012年第12期711-715,共5页Chinese Journal of Infectious Diseases

基　　金：十一五重大专项新发和突发传染病高通量检测技术的开发及其临床应用评估（2009ZXl0004-104）;上海市重点学科建设项目（B118）

摘　　要：目的比较犬。肾细胞系（MDCK）、人喉表皮癌细胞（Hep-2）和非洲绿猴肾细胞（Vero）制备的MHV混合细胞与相应单一敏感细胞株对流行性感冒（流感）病毒、肠道病毒的分离结果。方法流感样患者咽拭子标本接种MHV混合细胞和MDCK细胞，手足口病患儿咽拭子或粪标本接种MHV混合细胞和Veto细胞，观察病毒致细胞病变效应（CPE），采用多重反转录一聚合酶链反应（mRT-PCR）检测甲、乙型流感病毒和肠道病毒。结果MDCK和Vero细胞的CPE较MHV混合细胞明显。138份流感病毒接种MHV混合细胞，分离出34株流感病毒，分离率为24．6％；MDCK细胞分离出流感病毒39株，分离率为28．3％。MHV混合细胞与MDCK细胞流感病毒分离率比较，差异无统计学意义（X2=1．92，P〉0．05）。32株肠道病毒接种MHV混合细胞，分离出肠道病毒9株，分离率为28．1％；Vero细胞分离出12株肠道病毒，分离率为37．5％。MHV混合细胞与Vero细胞肠道病毒分离率比较，差异无统计学意义（X2=3．00，P〉0．05）。结论MHV混合细胞CPE不如单一细胞易于观察；MHV混合细胞适用于生物性突发公共卫生事件中，可分离培养临床标本中甲型、乙型流感病毒和表现为呼吸道症状的肠道病毒。Objective To evaluate shell vials of MHV, a combination of Madin-Darby canine kidney cells （MDCK）, human epidermoid cancer cells （Hep-2） and African green monkey kidney cells （Vero）, and conventional cell culture in detecting influenza viruses and enterovirus from fresh clinical specimens. Methods Specimens from patients with influenza-like illness and children with hand-foot- mouth disease were inoculated with both shell vials of MHV and MDCK/Vero. Then cytopathological effect （CPE） was examined daily. Influenza viruses and enteroviruses were detected by multiple reverse transcriptase-polymerase chain reaction （mRT-PCR）. Results CPE of MDCK/Vero cells were stronger than the shell vials of MHV. The isolation rate of influenza virus by MHV was 24.6 % （34/ 138） and that by MDCK was 28.3% （39/138）, which was not significantly different （X2=1.92, P〉0.05）. That of enterovirus by MHV was 28. 1% （9/32） and that by Vero was 37. 5%（12/32）, which was not significantly different （X2 =3.00, P〉0.05）. Conclusions CPE in MDCK/Vero cellsare easier to be observed than the shell vials of MHV. However, the shell vials of MHV are appropriate in public health emergencies, which can be used for isolation of in{luenza viruses and entm＇ovirus in patients with respiratory symptoms.

关 键 词：细胞 培养的 正黏病毒科 肠道病毒 细胞分离 

分 类 号：R4[医药卫生—临床医学] R51