禽流感病毒流式微球量子点探针免疫诊断新方法  

A Novel Immunoassay of Detecting Avian Influenza Virus by Cytometry Using Quantum Dots Conjugation Probe and Flowing Microsphere Carrier

作　　者：王楠[1,2] 邹明强[2] 汪明[1] 李锦丰[2] 薛强[2] 张帆[2] 

机构地区：[1]中国农业大学动物医学院,北京100193 [2]中国检验检疫科学研究院,北京100123

出　　处：《分析测试学报》2009年第7期764-768,共5页Journal of Instrumental Analysis

基　　金："十一五"国家科技支撑计划资助项目(2006BAK10B09);国家自然科学基金资助项目(20775076/B0509;20875084/B0509)

摘　　要：采用微波法水相中合成羧基化的绿色量子点,通过羧基与禽流感单克隆抗体氨基的共价结合,制备了检测禽流感病毒的探针,并结合流式微球技术,建立了量子点生物探针流式微球免疫检测禽流感病毒的新方法。以聚苯乙烯微球为蛋白质载体,将多克隆抗体包被到荧光微球上,依次加入待测抗原和量子点生物探针,形成双抗体夹心复合物,用流式细胞仪进行检测。实验结果表明,多抗和单抗的最佳质量浓度分别为92和4 mg/L,检测禽流感病毒比双抗体夹心ELISA灵敏16倍,比FITC标记单抗检测方法灵敏4倍。对阳性尿囊液的检测与ELISA呈现良好的相关性,不与鸡传染性支气管炎病毒、鸡马力克氏病毒、新城疫病毒等发生交叉反应。An aqueous solution of colloidal CdTe quantum dots（QDs） was prepared in the presence of MPA as a stabilizer. And QDs was used as fluorescent molecular probe after conjugating with a monoclonal antibody of avian influenza virus （AIV）. An immunoassay was developed for determination of AIV by cytometry with flowing fluorescence-coded microsphere as protein carrier. Polyclonal antibody （PcAb） was coupling with microsphere, then antigen and DQs labeled McAb（McAb- Qds）were added sequencely to form the sandwich of double antibodies, and the detection was performed by flow cytometry. PcAb concentration of 92 mg/L and McAb concentration of 4 mg/L were selected as the optimization working concentrations. Under the optimal conditions, the results indicated that, comparing with traditional methods, the proposed method was 16 times more sensitive than ELISA and 4 times than FITC conjugated method. The method showed well relative with the ELISA method, and no cross-reaction occured with infectious bronchitis virus （IBV）, marek＇s disease virus （MDV）and Newcastle disease virus （NDV）.

关 键 词：量子点探针 流式微球 禽流感病毒 免疫诊断 

分 类 号：R730.45[医药卫生—肿瘤] R373[医药卫生—临床医学]