title: Paper-based enzyme-free immunoassay for rapid detection and subtyping of influenza A H1N1 and H3N2 viruses.
abstract: Development of rapid screening in the ambulatory environment is the most pressing needs for the control of spread of infectious disease. Despite there are many methods to detect the immunoassay results, quantitative measurement in rapid disease screening is still a great challenge for point-of-care applications. In this work, based on the internal structural protein, i.e., nucleoprotein (NP), and outer surface glycoproteins, i.e., H1 and H3, of the influenza viruses, specific and sensitive immunoassay on paper-based platform was evaluated and confirmed. Detection and subtyping of influenza A H1N1 and H3N2 viruses found in people were demonstrated by colorimetric paper-based sandwich immunoassay. Concentration-dependent response to influenza viruses was shown and the detection limits could achieve 2.7×10(3) pfu/assay for H1 detection and 2.7×10(4) pfu/assay for H3 detection, which are within the clinical relevant level. Moreover, detection of influenza virus from infected cell lysate and clinical samples was demonstrated to further confirm the reliability of the paper-based immunoassay. The use of paper for the development of diagnostic devices has the advantages of lightweight, ease-of-use, and low cost and paper-based immunoassay is appropriate to apply for rapid screening in point-of-care applications.
authors.affiliation: Graduate Institute of Medical Mechatronics, College of Engineering, Chang Gung University, Taiwan; Department of Mechanical Engineering, College of Engineering, Chang Gung University, Taiwan. Electronic address: kflei@mail.cgu.edu.tw.
authors.forename: Kin Fong
authors.initials: KF
authors.lastname: Lei
authors.affiliation: Graduate Institute of Medical Mechatronics, College of Engineering, Chang Gung University, Taiwan.
authors.forename: Chia-Hao
authors.initials: CH
authors.lastname: Huang
authors.affiliation: Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Taiwan; Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taiwan; Department of Medical Biotechnology and Laboratory Science, College of Medicine, Chang Gung University, Taiwan. Electronic address: rlkuo@mail.cgu.edu.tw.
authors.forename: Rei-Lin
authors.initials: RL
authors.lastname: Kuo
authors.affiliation: Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Taiwan; Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taiwan.
authors.forename: Cheng-Kai
authors.initials: CK
authors.lastname: Chang
authors.affiliation: Department of Emergency Medicine, Chang Gung Memorial Hospital, Keelung, Taiwan; Clinical Informatics and Medical Statistics Research Center, Chang Gung University, Taiwan.
authors.forename: Kuan-Fu
authors.initials: KF
authors.lastname: Chen
authors.affiliation: Department of Medical Biotechnology and Laboratory Science, College of Medicine, Chang Gung University, Taiwan; Department of Laboratory Medicine, Chang Gung Memorial Hospital, Linkou, Taiwan.
authors.forename: Kuo-Chien
authors.initials: KC
authors.lastname: Tsao
authors.affiliation: School of Traditional Chinese Medicine, College of Medicine, Chang Gung University, Taiwan; Department of Radiation Oncology, Chang Gung Memorial Hospital, Linkou, Taiwan.
authors.forename: Ngan-Ming
authors.initials: NM
authors.lastname: Tsang
mesh_terms: D000818:Animals; D002460:Cell Line; D003124:Colorimetry / Q000295:instrumentation; D006801:Humans; D007118:Immunoassay / Q000295:instrumentation*; D053118:Influenza A Virus, H1N1 Subtype / Q000302:isolation & purification*; D053122:Influenza A Virus, H3N2 Subtype / Q000302:isolation & purification*; D007251:Influenza, Human / Q000175:diagnosis* / Q000821:virology; D057230:Limit of Detection; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation; D010209:Paper; D015203:Reproducibility of Results
keywords: Immunoassay; Influenza virus; Paper-based microfluidics