Gao Peng, Xu Guowang, Shi Xianzhe, Yuan Kailong, Tian Jing
National Chromatographic R. & A. Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China.
Electrophoresis. 2006 May;27(9):1784-9. doi: 10.1002/elps.200500656.
The rapid detection of pathogenic bacteria is extremely important in biotechnology and clinical diagnosis. CE has been utilized in the field of bacterial analysis for many years, but to some extent, simultaneous separation and identification of certain microbes from complex samples by CE coupled with UV detector is still a challenge. In this paper, we propose a new strategy for rapid separation and identification of Staphylococcus aureus (S. aureus) in bacterial mixtures by means of specific mAb-coated latex coupled with CZE. An appropriate set of conditions that selectively isolated S. aureus from the microorganisms Escherichia coli, Pseudomonas aeruginosa, and Klebsiella pneumoniae were established. S. aureus could be differentiated from the others by unique peaks in the electropherograms. The validity was also confirmed by LIF with antibodies specific to both the latex and the microbial cells. The LOD is as low as 9.0 x 10(5) colony forming unit/mL. We have also utilized this technology to identify S. aureus in a stool sample coming from a healthy volunteer spiked successfully with S. aureus. This CZE-UV technique can be applied to rapid diagnosis of enteritis caused by S. aureus or other bacterial control-related fields needing rapid identification of target pathogens from microbial mixtures. In theory, this method is suitable for the detection of any bacterium as long as corresponding bacterium-specific antibody-coated latex is available.
病原菌的快速检测在生物技术和临床诊断中极为重要。毛细管电泳(CE)已在细菌分析领域应用多年,但在一定程度上,利用CE结合紫外检测器从复杂样品中同时分离和鉴定某些微生物仍是一项挑战。本文提出了一种新策略,通过特异性单克隆抗体包被的乳胶与毛细管区带电泳(CZE)联用,快速分离和鉴定细菌混合物中的金黄色葡萄球菌(S. aureus)。建立了一组适当的条件,可从大肠杆菌、铜绿假单胞菌和肺炎克雷伯菌中选择性分离出金黄色葡萄球菌。金黄色葡萄球菌可通过电泳图中的独特峰与其他细菌区分开来。通过使用针对乳胶和微生物细胞的特异性抗体的激光诱导荧光(LIF)也证实了其有效性。检测限低至9.0×10⁵菌落形成单位/毫升。我们还利用该技术鉴定了来自一名成功接种金黄色葡萄球菌的健康志愿者的粪便样本中的金黄色葡萄球菌。这种CZE-UV技术可应用于金黄色葡萄球菌引起的肠炎的快速诊断或其他需要从微生物混合物中快速鉴定目标病原体的细菌控制相关领域。理论上,只要有相应的细菌特异性抗体包被的乳胶,该方法适用于任何细菌的检测。