压电免疫传感器微阵列检测铜绿假单胞菌的实验研究

Detection of Pseudomonas aeruginosa by Piezoelectric Immunosensor Microarray: An Experimental Research

目的 构建一种新型压电铜绿假单胞菌免疫传感器, 并对铜绿假单胞菌进行检测,以探索临床应用的可行性。方法 在免疫传感器微阵列表面固定铜绿假单胞菌单克隆抗体,对铜绿假单胞菌菌液进行检测,绘制用于检测的标准曲线并进行曲线拟合,并对50例临床标本进行检测,结果与传统分析方法进行对比。结果 铜绿假单胞菌菌液的浓度在103~109细胞/ml范围内,菌量与传感器频率的变化之间呈良好的指数线性关系,指数式回归方程为△F =12.6625×C0 1757,r=0.9949;通过对未知浓度的铜绿假单胞菌菌液进行检测,定量结果与吖啶橙荧光显微镜直接计数法(AODC)的相关系数r为0.973;50 例临床标本检测结果与常规培养检测法高度符合(P>0.05)。结论 应用压电石英晶体铜绿假单胞菌微阵列免疫传感器能够对铜绿假单胞菌进行的定量和定性分析,该技术应用于检测铜绿假单胞菌具有时间短、结果准确、无需长时间培养等特点,有较高的推广应用价值。

OBJECTIVE Based on the construction of piezoelectric immunosensor microarray detection system, to establish a new method to detect Pseudomonas aeruginosa (PA), and to explore the possibility of clinical application. METHODS Monoclonal antibody for detecting PA was immobilized on the gold electrode surface of immunosensor microarray. And the quantitative curve was statistically drawn up about the relationship between the concentration of PA and decrease in frequency, and to explore the possibility of the clinical application, 50 clinical samples were detected by our immunosensor microarray, the results were compared those with conventional experiments. RESULTS The statistic exponential regression equation was △F =12.6625×C^(0.1757) among the range of PA concentration from 10~3 to 10~9 cells/ml and the correlative coefficient was 0.9949. When unknown concentration of PA was detected using immunosensor microarray, and the results were compared with those of AODC, the correlation coefficient was 0.937. The analytical results of 50 clinical samples given by this approach were in satisfactory agreement with those given by conventional experiments(P>0.05). CONCLUSIONS Quantitative and qualitative analysis of PA can be proceeded by immunosensor microarray. Piezoelectric immunosensor microarray is a new accurate method for detecting and identifying PA, with less time consumed, and hence has great value in clinical practice.