CAJ | 학술논문

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.
채용미파법수상중합성최기화적록색양자점,통과최기여금류감단극륭항체안기적공개결합,제비료검측금류감병독적탐침,병결합류식미구기술,건립료양자점생물탐침류식미구면역검측금류감병독적신방법.이취분을희미구위단백질재체,장다극륭항체포피도형광미구상,의차가입대측항원화양자점생물탐침,형성쌍항체협심복합물,용류식세포의진행검측.실험결과표명,다항화단항적최가질량농도분별위92화4 mg/L,검측금류감병독비쌍항체협심ELISA령민16배,비FITC표기단항검측방법령민4배.대양성뇨낭액적검측여ELISA정현량호적상관성,불여계전염성지기관염병독、계마력극씨병독、신성역병독등발생교차반응.
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).