title: Centrifugal loop-mediated isothermal amplification microdevice for rapid, multiplex and colorimetric foodborne pathogen detection.
abstract: We present a centrifugal microfluidic device which enables multiplex foodborne pathogen identification by loop-mediated isothermal amplification (LAMP) and colorimetric detection using Eriochrome Black T (EBT). Five identical structures were designed in the centrifugal microfluidic system to perform the genetic analysis of 25 pathogen samples in a high-throughput manner. The sequential loading and aliquoting of the LAMP cocktail, the primer mixtures, and the DNA sample solutions were accomplished by the optimized zigzag-shaped microchannels and RPM control. We targeted three kinds of pathogenic bacteria (Escherichia coli O157:H7, Salmonella typhimurium and Vibrio parahaemolyticus) and detected the amplicons of the LAMP reaction by the EBT-mediated colorimetric method. For the limit-of-detection (LOD) test, we carried out the LAMP reaction on a chip with serially diluted DNA templates of E. coli O157:H7, and could observe the color change with 380 copies. The used primer sets in the LAMP reaction were specific only to the genomic DNA of E. coli O157:H7, enabling the on-chip selective, sensitive, and high-throughput pathogen identification with the naked eyes. The entire process was completed in 60min. Since the proposed microsystem does not require any bulky and expensive instrumentation for end-point detection, our microdevice would be adequate for point-of-care (POC) testing with high simplicity and high speed, providing an advanced genetic analysis microsystem for foodborne pathogen detection.
authors: Department of Chemical and Biomolecular Engineering (BK21 PLUS Program), Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors: Seung Jun
authors: SJ
authors: Oh
authors: Department of Chemical and Biomolecular Engineering (BK21 PLUS Program), Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors: Byung Hyun
authors: BH
authors: Park
authors: Department of Chemical and Biomolecular Engineering (BK21 PLUS Program), Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors: Jae Hwan
authors: JH
authors: Jung
authors: Department of Chemical and Biomolecular Engineering (BK21 PLUS Program), Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors: Goro
authors: G
authors: Choi
authors: Department of Chemical and Biomolecular Engineering (BK21 PLUS Program), Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors: Doh C
authors: DC
authors: Lee
authors: Department of Chemical and Biomolecular Engineering (BK21 PLUS Program), Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors: Do Hyun
authors: DH
authors: Kim
authors: Department of Chemical and Biomolecular Engineering (BK21 PLUS Program), Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea. Electronic address: seots@kaist.ac.kr.
authors: Tae Seok
authors: TS
authors: Seo
mesh_terms: D015374:Biosensing Techniques; D003124:Colorimetry; D004269:DNA, Bacterial / Q000302:isolation & purification*; D019453:Escherichia coli O157 / Q000302:isolation & purification / Q000472:pathogenicity; D005517:Foodborne Diseases / Q000175:diagnosis / Q000382:microbiology*; D006801:Humans; D056656:Lab-On-A-Chip Devices; D057230:Limit of Detection; D012486:Salmonella typhimurium / Q000302:isolation & purification / Q000472:pathogenicity; D014736:Vibrio parahaemolyticus / Q000302:isolation & purification / Q000472:pathogenicity
chemical_list: D004269:DNA, Bacterial
keywords: Centrifugal microdevice; Colorimetric detection; Foodborne pathogen; Loop-mediated isothermal amplification; Multiplex detection