title: An integrated microfluidic PCR system with immunomagnetic nanoparticles for the detection of bacterial pathogens.
abstract: There is growing interest in rapid microbial pre-concentration methods to lower the detection limit of bacterial pathogens of low abundance in samples. Here, we report an integrated microfluidic PCR system that enables bacterial cells of interest in samples to be concentrated prior to PCR. It consists of two major compartments: a preconcentration chamber for the immunomagnetic separation of bacterial cells, and a PCR chamber for the DNA amplification of the concentrated cells. We demonstrate the feasibility of the system for the detection of microbial pathogens by preconcentrating the human pathogen Escherichia coli O157:H7, and also amplifying its DNA. The detection limit of E. coli O157:H7 in the PCR system is 1 × 103CFU (colony forming unit)/mL. On-chip processing steps, including preconcentration and PCR steps, take less than two hours. Our system can serve as a rapid, specific, and quantitative platform for the detection of microbial pathogens in samples of large volume.
authors.affiliation: School of Mechanical Engineering, Sungkyunkwan University, Suwon, 16419, Korea.
authors.forename: Irisappan
authors.initials: I
authors.lastname: Ganesh
authors.affiliation: Department of BioNano Technology, College of BioNano Technology, Gachon University, Seongnam, Gyeonggi-do, 13120, Korea.
authors.forename: Buu Minh
authors.initials: BM
authors.lastname: Tran
authors.affiliation: School of Mechanical Engineering, Sungkyunkwan University, Suwon, 16419, Korea.
authors.forename: Yonghee
authors.initials: Y
authors.lastname: Kim
authors.affiliation: School of Mechanical Engineering, Sungkyunkwan University, Suwon, 16419, Korea.
authors.forename: Jaewon
authors.initials: J
authors.lastname: Kim
authors.affiliation: Department of Chemistry and Nano Sciences (BK21 plus), Ewha Womans University, Seoul, 03760, Korea.
authors.forename: Hua
authors.initials: H
authors.lastname: Cheng
authors.affiliation: Department of BioNano Technology, College of BioNano Technology, Gachon University, Seongnam, Gyeonggi-do, 13120, Korea. nylee@gachon.ac.kr.
authors.forename: Nae Yoon
authors.initials: NY
authors.lastname: Lee
authors.affiliation: School of Mechanical Engineering, Sungkyunkwan University, Suwon, 16419, Korea. nanopark@skku.edu.
authors.forename: Sungsu
authors.identifier: 0000-0003-3062-1302
authors.initials: S
authors.lastname: Park
mesh_terms: D004269:DNA, Bacterial / Q000235:genetics; D004867:Equipment Design; D019453:Escherichia coli O157 / Q000166:cytology / Q000235:genetics* / Q000302:isolation & purification*; D005290:Ferric Compounds / Q000737:chemistry*; D023281:Genomics; D018189:Immunomagnetic Separation / Q000295:instrumentation*; D056656:Lab-On-A-Chip Devices; D053758:Nanoparticles; D016133:Polymerase Chain Reaction / Q000295:instrumentation*; D018511:Systems Integration; D013696:Temperature
chemical_list: D004269:DNA, Bacterial; D005290:Ferric Compounds; C000499:ferric oxide
keywords: Escherichia coli O157:H7; Immunomagnetic separation; Microfluidic device; On-chip PCR