title: A novel sensitive pathogen detection system based on Microbead Quantum Dot System.
abstract: A fast and accurate detection system for pathogens can provide immediate measurements for the identification of infectious agents. Therefore, the Microbead Quantum-dots Detection System (MQDS) was developed to identify and measure target DNAs of pathogenic microorganisms and eliminated the need of PCR amplifications. This nanomaterial-based technique can detect different microorganisms by flow cytometry measurements. In MQDS, pathogen specific DNA probes were designed to form a hairpin structure and conjugated on microbeads. In the presence of the complementary target DNA sequence, the probes will compete for binding with the reporter probes but will not interfere with the binding between the probe and internal control DNA. To monitor the binding process by flow cytometry, both the reporter probes and internal control probes were conjugated with Quantum dots that fluoresce at different emission wavelengths using the click reaction. When MQDS was used to detect the pathogens in environmental samples, a high correlation coefficient (R=0.994) for Legionella spp., with a detection limit of 0.1 ng of the extracted DNAs and 10 CFU/test, can be achieved. Thus, this newly developed technique can also be applied to detect other pathogens, particularly viruses and other genetic diseases.
authors.affiliation: Department of Bioscience Technology, Chung Yuan Christian University, Chung-Li 320, Taiwan; Center for Nanotechnology and Center for Biomedical Technology, Chung Yuan Christian University, Chung-Li 320, Taiwan; R&D Center of Membrane Technology, Chung Yuan Christian University, Chung-Li 320, Taiwan.
authors.forename: Tzong-Yuan
authors.initials: TY
authors.lastname: Wu
authors.affiliation: Department of Bioscience Technology, Chung Yuan Christian University, Chung-Li 320, Taiwan.
authors.forename: Yi-Yu
authors.initials: YY
authors.lastname: Su
authors.affiliation: Department of Bioscience Technology, Chung Yuan Christian University, Chung-Li 320, Taiwan.
authors.forename: Wei-Hsien
authors.initials: WH
authors.lastname: Shu
authors.affiliation: Department of Bioscience Technology, Chung Yuan Christian University, Chung-Li 320, Taiwan; Department of Chemistry, Chung Yuan Christian University, Chung-Li 320, Taiwan.
authors.forename: Augustus T
authors.initials: AT
authors.lastname: Mercado
authors.affiliation: Environmental Analysis Laboratory, Environmental Protection Administration, Chung-Li 320, Taiwan.
authors.forename: Shi-Kwun
authors.initials: SK
authors.lastname: Wang
authors.affiliation: Environmental Analysis Laboratory, Environmental Protection Administration, Chung-Li 320, Taiwan.
authors.forename: Ling-Yi
authors.initials: LY
authors.lastname: Hsu
authors.affiliation: Department of Chemistry, Chung Yuan Christian University, Chung-Li 320, Taiwan.
authors.forename: Yow-Fu
authors.initials: YF
authors.lastname: Tsai
authors.affiliation: Department of Bioscience Technology, Chung Yuan Christian University, Chung-Li 320, Taiwan; Center for Nanotechnology and Center for Biomedical Technology, Chung Yuan Christian University, Chung-Li 320, Taiwan. Electronic address: cychen@cycu.edu.tw.
authors.forename: Chung-Yung
authors.initials: CY
authors.lastname: Chen
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D057930:Click Chemistry; D004269:DNA, Bacterial / Q000737:chemistry / Q000302:isolation & purification*; D005434:Flow Cytometry; D005453:Fluorescence; D006801:Humans; D007875:Legionella / Q000302:isolation & purification* / Q000472:pathogenicity; D057230:Limit of Detection; D008863:Microspheres; D045663:Quantum Dots / Q000737:chemistry*
chemical_list: D004269:DNA, Bacterial
keywords: Flow cytometry; Hairpin structure; Microbead Quantum-dots Detection System (MQDS); Pathogen; Quantum dots; The click reaction