荧光量子点免疫标记法检测炭疽芽孢杆菌

Detection of Bacillus Anthracis Using Fluorescence Immunoassay with Quantum Dots Labels

建立了荧光量子点标记-免疫分析技术联用检测炭疽芽孢杆菌的方法。通过抗原抗体反应,结合生物素与亲和素间的特异性相互作用,将QDs特异性标记在炭疽芽孢杆菌上,并利用荧光显微镜和荧光分光光度计进行了验证。采用实验室自制的便携式荧光检测系统对标记QDs的炭疽芽孢杆菌样品进行定量检测。结果表明,在炭疽芽孢杆菌浓度在100～1×106 CFU/mL范围内,相对荧光强度与炭疽芽孢杆菌浓度呈良好的线性关系,相关系数R=0.9554,检测相对标准偏差为2.2%。通过与同菌属其它杆菌对比,证明本方法特异性良好。与传统方法相比,本方法操作简单,检测时间短(1 h),且能实现定量检测,在公共安全等方面有广泛的应用前景。

A method for detecting Bacillus anthracis（B.anthracis） was constructed with indirect immunofluorescence technique using quantum dots（QDs） as fluorescent probes.B.anthracis could be labeled with QDs selectively,that was proved by fluorescence microscope and F4500 spectrofluoro-meter.The laboratory-made system including biosensor and device was utilized as detection apparatus.A linear relationship of the fluorescence intensity and log total count of B.anthracis was obtained with correlation coefficient of 0.9554 from 100 CFU/mL to 1×10^6 CFU/mL,and a low relative standard deviation of 2.2%.By comparing the fluorescence intensity of B.anthracis and other bacillus,this method showed good specificity.Because of the simple operation,the detecting time is less（1 h） than traditional methods as well as quantitative detection,this method has potential prospect in monitoring B.anthracis and other biowarfare agents.