禽流感病毒流式微球量子点探针免疫诊断新方法

摘要

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 virus(NDV).%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.