Real-Time PCR与流感病毒分离关系研究  

The research of relationship between Real-Time PCR and influenza virus isolation

作　　者：刘磊[1] 王彦玲[1] 吕昌武[1] 吴双志[1] 孙宇[1] 谭君[1] 沙丽娜[1] 

机构地区：[1]牡丹江市疾病预防控制中心,黑龙江157021

出　　处：《医学动物防制》2017年第5期580-582,共3页Journal of Medical Pest Control

基　　金：牡丹江市科学技术计划项目-牡丹江市流感优势毒株的监测及流感防控策略研究(Z2015S0022)

摘　　要：目的研究提高流感病毒分离率的方法。方法 Real-Time PCR检测500份流感监测病例,选择甲型流感H1N1、H3N2和乙型流感阳性及阴性样品各10份共计40份,分别接种MDCK细胞和鸡胚,用红血球凝集试验测定病毒滴度,以血凝抑制试验鉴定毒株型别。结果阳性样品中,MDCK细胞分离出季节性甲型流感H1N1型8份、H3N2型7份、乙型流感8份;鸡胚分离出季节性甲型流感H1N1型3份、H3N2型0份、乙型流感5份。阴性样品中,MDCK细胞分离出季节性甲型流感H1N1型3份、H3N2型3份;鸡胚分离出季节性甲型流感H1N1型2份、H3N2型0份。结论 MDCK细胞分离流感病毒时,Ct值已经在阴性范围之内的样品也应进行病毒分离,以提高病毒分离率;鸡胚分离流感病毒时,应选择MDCK细胞接毒一代且滴度达到1∶128的强毒株进行分离。Objective To investigate the methods for improving the isolating rate of influenza virus. Methods Real Time PCR have detected 500 influenza monitoring cases, and 10 specimens positive and 10 specimens negative separately from the Vacci- hum Influenzae Vivum H1N1, H3N2 and Influenza B were choosed to be separately inoculated on MDCK cell culture, and into chick- embryo allantoic and amniotic cavity. Virus titers were detected by hemagglutination tests（HA） and subtyped by hemag- glutination inhibition tests（HI）. Results In specimens positive, 8 H1N1, 7 H3N2 and 8 Influenza B seasonality influenza virus were obtained by MDCK cell culture, and 3 H1N1, 0 H3N2 and 5 Influenza B by the chick - embryo technics. In specimens negative, 3 H1N1 and 3 H3N2 seasonality influenza virus were obtained by MDCK cell culture, and 2 H1N1 and 0 H3N2 by the chick -embryo technics. Conclusion Isolating influenza virus by MDCK cell culture, the specimens that Ct was already in the negative range should isolate virus to improve the rate of virus isolation. When isolating influenza virus by chick - embryo inoculation, we should choose one generation of the MDCK cell culture that the virus titers achieve 1：128.

关 键 词：流感病毒 病毒分离 细胞培养 鸡胚接种 

分 类 号：R373.13[医药卫生—病原生物学]