title: An integrated rotary microfluidic system with DNA extraction, loop-mediated isothermal amplification, and lateral flow strip based detection for point-of-care pathogen diagnostics.
abstract: Point-of-care (POC) molecular diagnostics plays a pivotal role for the prevention and treatment of infectious diseases. In spite of recent advancement in microfluidic based POC devices, there are still rooms for development to realize rapid, automatic and cost-effective sample-to-result genetic analysis. In this study, we propose an integrated rotary microfluidic system that is capable of performing glass microbead based DNA extraction, loop mediated isothermal amplification (LAMP), and colorimetric lateral flow strip based detection in a sequential manner with an optimized microfluidic design and a rotational speed control. Rotation direction-dependent coriolis force and siphon valving structures enable us to perform the fluidic control and metering, and the use of the lateral flow strip as a detection method renders all the analytical processes for nucleic acid test simplified and integrated without the need of expensive instruments or human intervention. As a proof of concept for point-of-care DNA diagnostics, we identified the food-borne bacterial pathogen which was contaminated in water or milk. Not only monoplex Salmonella Typhimurium but also multiplex Salmonella Typhimurium and Vibrio parahaemolyticus were analysed on the integrated rotary genetic analysis microsystem with a limit of detection of 50 CFU in 80min. In addition, three multiple samples were simultaneously analysed on a single device. The sample-to-result capability of the proposed microdevice provides great usefulness in the fields of clinical diagnostics, food safety and environment monitoring.
authors.affiliation: Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Byung Hyun
authors.initials: BH
authors.lastname: Park
authors.affiliation: Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Seung Jun
authors.initials: SJ
authors.lastname: Oh
authors.affiliation: Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Jae Hwan
authors.initials: JH
authors.lastname: Jung
authors.affiliation: Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Goro
authors.initials: G
authors.lastname: Choi
authors.affiliation: Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Ji Hyun
authors.initials: JH
authors.lastname: Seo
authors.affiliation: Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Do Hyun
authors.initials: DH
authors.lastname: Kim
authors.affiliation: Department of Chemical Engineering, College of Engineering, Kyung Hee University, 1 Seochon-dong, Giheung-gu, Yongin-si, Gyeonggi-do 17140, Republic of Korea.
authors.forename: Eun Yeol
authors.initials: EY
authors.lastname: Lee
authors.affiliation: Department of Chemical Engineering, College of Engineering, Kyung Hee University, 1 Seochon-dong, Giheung-gu, Yongin-si, Gyeonggi-do 17140, Republic of Korea. Electronic address: seots@khu.ac.kr.
authors.forename: Tae Seok
authors.initials: TS
authors.lastname: Seo
mesh_terms: D000818:Animals; D015374:Biosensing Techniques / Q000295:instrumentation; D003124:Colorimetry / Q000295:instrumentation; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics / Q000302:isolation & purification*; D004867:Equipment Design; D006801:Humans; D056656:Lab-On-A-Chip Devices; D057230:Limit of Detection; D008892:Milk / Q000382:microbiology*; D021141:Nucleic Acid Amplification Techniques / Q000295:instrumentation; D019095:Point-of-Care Systems; D011934:Reagent Strips / Q000032:analysis; D012480:Salmonella Infections / Q000175:diagnosis / Q000382:microbiology; D012486:Salmonella typhimurium / Q000235:genetics / Q000302:isolation & purification*; D052616:Solid Phase Extraction / Q000295:instrumentation*; D014735:Vibrio Infections / Q000175:diagnosis / Q000382:microbiology; D014736:Vibrio parahaemolyticus / Q000235:genetics / Q000302:isolation & purification*; D014871:Water Microbiology
chemical_list: D004269:DNA, Bacterial; D011934:Reagent Strips
keywords: Centrifugal microdevice; Lateral flow strip detection; Loop-mediated isothermal amplification; Point-of-care diagnostics; Solid-phase DNA extraction