一种快速诊断甲型H1N1流感病毒的悬浮芯片方法  

A METHOD FOR RAPID DIAGNOSIS OF A(H1N1) INFLUENZA VIRUS BY USING SUSPENSION ARRAY

作　　者：孙智勇[1] 宁保安[1] 牛超[1] 苏璞[1] 彭媛[1] 白家磊[1] 高志贤[1] 

机构地区：[1]军事医学科学院卫生学环境医学研究所、天津环境与食品安全风险监控技术重点实验室,天津300050

出　　处：《解放军预防医学杂志》2012年第3期172-175,共4页Journal of Preventive Medicine of Chinese People's Liberation Army

基　　金："十二五"农村领域国家科技计划课题(No.2012AA101604)

摘　　要：目的建立一种可快速诊断甲型H1N1流感病毒的悬浮芯片方法,为研制快速检测H1N1流感病毒试剂盒奠定基础。方法针对HIN1特有的NA基因、HA基因、NS基因序列设计引物,在下游引物末端用生物素标记。根据3个基因扩增区内的核酸序列设计特异性探针,探针末端修饰氨基C12,以便把特异性的核酸探针共价结合到用彩色荧光编码的微球表面。采用液态悬浮杂交方法,将微球与生物素标记的目标物进行杂交,通过结合的链霉亲和素藻红蛋白,检测目标物。结果所设计的探针没有交叉反应,单通道检测探针灵敏度HA基因为1.6 nmol/L,NA基因、NS基因均为8.0 nmol/L;多通道检测的探针灵敏度为1.6 nmol/L。结论该方法准确、灵敏度高,可以用于下一步悬浮芯片检测试剂盒的研制。Objective To provide a method for rapid diagnosis of A （H1N1） influenza virus by suspension chip. Method According to the NA, HA, and NS gene sequences, primers were designed for the high specificity of the H1N1 influenza virus. The terminal of the downstream primer was labeled with biotin. Based on the three genes amplifica- tion sequences of nucleic acids, specific probe was designed, and the probe end was modified with NH2-C12. The Carboxy- lated coded microspheres were coupled with the probe by the treatment with EDC solution. Then the microspheres were hy- bridized with target labeled with biotin through suspension hybridization method. The target was detected through the combination of streptavidin-R-phycoerythrin （SA-PE）. Results The designed probe was without cross-reaction. In single chan- nel test, the minimum detection sensitivity for HA gene was 1.6 nmol/L, for both NA, and NS gene 8 nmol/L; in muhichannel test, the minimum detection sensitivity was 1.6 nmol/L. Conclusion The method, which is accurate and sensitive, is suitable to be used for further developing suspension chip testing kit.

关 键 词：H1N1 快速诊断 悬浮芯片 

分 类 号：R373.13[医药卫生—病原生物学]