title: Ultrasensitive detection of bacteria by microchip electrophoresis based on multiple-concentration approaches combining chitosan sweeping, field-amplified sample stacking, and reversed-field stacking.
abstract: In this paper we describe an on-chip multiple-concentration method combining chitosan (CS) sweeping, reversed-field stacking, and field-amplified sample stacking for highly efficient detection of bacteria. Escherichia coli was selected as a model bacterium to investigate the efficiency of this multiple-concentration method. CS was the most suitable sweeping agent for microchip electrophoresis, replacing the usually used cetyltrimethylammonium bromide for capillary electrophoresis. The additive taurine had a synergistic effect by enhancing the interaction between CS and the surface of the bacteria, thus improving the analysis sensitivity. All steps of the concentration method and related mechanisms are described and discussed in detail. A concentration enhancement factor of approximately 6000 was obtained using this concentration method under optimal conditions as compared to using no concentration step, and the detection limit of E. coli was 145 CFU/mL. The multiple-concentration methodology was also applied for the quantification of bacteria in surface water, and satisfactory results were achieved. The application of this methodology showed that the concentration enhancement of bacteria clearly conferred advantageous sensitivity, speed, and sample volume compared to established methods.
authors.affiliation: Department of Chemistry, East China Normal University, Shanghai 200062, People's Republic of China.
authors.forename: Zhi-Fang
authors.initials: ZF
authors.lastname: Wang
authors.forename: Shuang
authors.initials: S
authors.lastname: Cheng
authors.forename: Shu-Li
authors.initials: SL
authors.lastname: Ge
authors.forename: Huan
authors.initials: H
authors.lastname: Wang
authors.forename: Qing-Jiang
authors.initials: QJ
authors.lastname: Wang
authors.forename: Pin-Gang
authors.initials: PG
authors.lastname: He
authors.forename: Yu-Zhi
authors.initials: YZ
authors.lastname: Fang
mesh_terms: D000077286:Cetrimonium; D002593:Cetrimonium Compounds / Q000737:chemistry; D048271:Chitosan / Q000737:chemistry*; D046209:Electrophoresis, Microchip; D004926:Escherichia coli / Q000302:isolation & purification*; D014871:Water Microbiology
chemical_list: D002593:Cetrimonium Compounds; D048271:Chitosan; D000077286:Cetrimonium