title: A fully disposable and integrated paper-based device for nucleic acid extraction, amplification and detection.
abstract: Nucleic acid testing (NAT) has been widely used for disease diagnosis, food safety control and environmental monitoring. At present, NAT mainly involves nucleic acid extraction, amplification and detection steps that heavily rely on large equipment and skilled workers, making the test expensive, time-consuming, and thus less suitable for point-of-care (POC) applications. With advances in paper-based microfluidic technologies, various integrated paper-based devices have recently been developed for NAT, which however require off-chip reagent storage, complex operation steps and equipment-dependent nucleic acid amplification, restricting their use for POC testing. To overcome these challenges, we demonstrate a fully disposable and integrated paper-based sample-in-answer-out device for NAT by integrating nucleic acid extraction, helicase-dependent isothermal amplification and lateral flow assay detection into one paper device. This simple device allows on-chip dried reagent storage and equipment-free nucleic acid amplification with simple operation steps, which could be performed by untrained users in remote settings. The proposed device consists of a sponge-based reservoir and a paper-based valve for nucleic acid extraction, an integrated battery, a PTC ultrathin heater, temperature control switch and on-chip dried enzyme mix storage for isothermal amplification, and a lateral flow test strip for naked-eye detection. It can sensitively detect Salmonella typhimurium, as a model target, with a detection limit of as low as 102CFU ml-1in wastewater and egg, and 103CFU ml-1in milk and juice in about an hour. This fully disposable and integrated paper-based device has great potential for future POC applications in resource-limited settings.
authors.affiliation: School of Life Sciences, Northwestern Polytechnical University, Xi'an 710072, P.R. China. kittyyh@nwpu.edu.cn and Key Laboratory for Space Bioscience and Biotechnology, Northwestern Polytechnical University, Xi'an 710072, P.R. China and Bioinspired Engineering and Biomechanics Center (BEBC), Xi'an Jiaotong University, Xi'an 710049, P.R. China.
authors.forename: Ruihua
authors.initials: R
authors.lastname: Tang
authors.affiliation: School of Life Sciences, Northwestern Polytechnical University, Xi'an 710072, P.R. China. kittyyh@nwpu.edu.cn and Key Laboratory for Space Bioscience and Biotechnology, Northwestern Polytechnical University, Xi'an 710072, P.R. China.
authors.forename: Hui
authors.initials: H
authors.lastname: Yang
authors.affiliation: The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an 710049, P.R. China. fengxu@mail.xjtu.edu.cn and Bioinspired Engineering and Biomechanics Center (BEBC), Xi'an Jiaotong University, Xi'an 710049, P.R. China and Xi'an Diandi Biotech Company, Xi'an 710049, P.R. China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Gong
authors.affiliation: The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an 710049, P.R. China. fengxu@mail.xjtu.edu.cn and Bioinspired Engineering and Biomechanics Center (BEBC), Xi'an Jiaotong University, Xi'an 710049, P.R. China and Xi'an Diandi Biotech Company, Xi'an 710049, P.R. China.
authors.forename: MinLi
authors.initials: M
authors.lastname: You
authors.affiliation: Bioinspired Engineering and Biomechanics Center (BEBC), Xi'an Jiaotong University, Xi'an 710049, P.R. China and Key Laboratory of Thermo-Fluid Science and Engineering of Ministry of Education, School of Energy and Power Engineering, Xi'an Jiaotong University, Xi'an 710049, P.R. China.
authors.forename: Zhi
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Bioinspired Engineering and Biomechanics Center (BEBC), Xi'an Jiaotong University, Xi'an 710049, P.R. China.
authors.forename: Jane Ru
authors.initials: JR
authors.lastname: Choi
authors.affiliation: Xi'an Diandi Biotech Company, Xi'an 710049, P.R. China.
authors.forename: Ting
authors.initials: T
authors.lastname: Wen
authors.affiliation: Key Laboratory of Thermo-Fluid Science and Engineering of Ministry of Education, School of Energy and Power Engineering, Xi'an Jiaotong University, Xi'an 710049, P.R. China.
authors.forename: Zhiguo
authors.identifier: 0000-0003-1469-6152
authors.initials: Z
authors.lastname: Qu
authors.affiliation: School of Life Sciences, Northwestern Polytechnical University, Xi'an 710072, P.R. China. kittyyh@nwpu.edu.cn and Key Laboratory for Space Bioscience and Biotechnology, Northwestern Polytechnical University, Xi'an 710072, P.R. China.
authors.forename: Qibing
authors.initials: Q
authors.lastname: Mei
authors.affiliation: The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an 710049, P.R. China. fengxu@mail.xjtu.edu.cn and Bioinspired Engineering and Biomechanics Center (BEBC), Xi'an Jiaotong University, Xi'an 710049, P.R. China.
authors.forename: Feng
authors.identifier: 0000-0003-4351-0222
authors.initials: F
authors.lastname: Xu
mesh_terms: D001628:Beverages / Q000382:microbiology; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics / Q000302:isolation & purification*; D004867:Equipment Design; D056656:Lab-On-A-Chip Devices; D057230:Limit of Detection; D021141:Nucleic Acid Amplification Techniques / Q000295:instrumentation*; D009696:Nucleic Acids / Q000032:analysis / Q000235:genetics / Q000302:isolation & purification; D010209:Paper; D012486:Salmonella typhimurium / Q000235:genetics
chemical_list: D004269:DNA, Bacterial; D009696:Nucleic Acids