Nakamura N, Burgess J G, Yagiuda K, Kudo S, Sakaguchi T, Matsunaga T
Department of Biotechnology, Tokyo University of Agriculture and Technology, Japan.
Anal Chem. 1993 Aug 1;65(15):2036-9. doi: 10.1021/ac00063a018.
A novel fluoroimmunoassay method using bacterial magnetic particles for the highly sensitive detection of bacteria has been developed. Fluorescein isothiocyanate (FITC) conjugated monoclonal anti-Escherichia coli antibody was immobilized onto bacterial magnetic particles (BMPs) using a heterobifunctional reagent, N-succinimdyl 3-(2-pyridyldithio)propionate (SPDP). E. coli cells were reacted with FITC-antibody-BMP conjugates for 15 min in an inhomogeneous magnetic field which enhanced aggregation. The cell/BMP complexes sedimented, causing relative fluorescence intensity of the solution to decrease with increasing microbial cell concentration. A linear relationship was obtained between the relative fluorescence intensity and cell concentration in the range of 10(2)-10(6) cells/mL. Selectivity of this detection system was satisfactory. Monoclonal antibody immobilized on BMPs was also applied to the specific removal of E. coli from the bacterial suspension.
一种利用细菌磁性颗粒进行细菌高灵敏度检测的新型荧光免疫分析方法已被开发出来。使用异双功能试剂N-琥珀酰亚胺基-3-(2-吡啶二硫代)丙酸酯（SPDP）将异硫氰酸荧光素（FITC）偶联的抗大肠杆菌单克隆抗体固定在细菌磁性颗粒（BMPs）上。大肠杆菌细胞在增强聚集的非均匀磁场中与FITC-抗体-BMP偶联物反应15分钟。细胞/BMP复合物沉淀，导致溶液的相对荧光强度随着微生物细胞浓度的增加而降低。在10(2)-10(6)个细胞/毫升的范围内，相对荧光强度与细胞浓度之间获得了线性关系。该检测系统的选择性令人满意。固定在BMPs上的单克隆抗体也被用于从细菌悬浮液中特异性去除大肠杆菌。
