title: Rapid and quantitative detection of zoonotic influenza A virus infection utilizing coumarin-derived dendrimer-based fluorescent immunochromatographic strip test (FICT).
abstract: Great efforts have been made to develop robust signal-generating fluorescence materials which will help in improving the rapid diagnostic test (RDT) in terms of sensitivity and quantification. In this study, we developed coumarin-derived dendrimer-based fluorescent immunochromatographic strip test (FICT) assay with enhanced sensitivity as a quantitative diagnostic tool in typical RDT environments. The accuracy of the proposed FICT was compared with that of dot blot immunoassay techniques and conventional RDTs. Through conjugation of coumarin-derived dendrimers with latex beads, fluorescent emission covering broad output spectral ranges was obtained which provided a distinct advantage of easy discrimination of the fluorescent emission of the latex beads with a simple insertion of a long-pass optical filter away from the excitation wavelength. The newly developed FICT assay was able to detect 100 ng/10 μL of influenza A nucleoprotein (NP) antigen within 5 minutes, which corresponded to 2.5-fold higher sensitivity than that of the dot blot immunoassay or conventional RDTs. Moreover, the FICT assay was confirmed to detect at least four avian influenza A subtypes (H5N3, H7N1, H7N7, and H9N2). On applying the FICT to the clinical swab samples infected with respiratory viruses, our FICT assay was confirmed to differentiate influenza H1N1 infection from other respiratory viral diseases. These data demonstrate that the proposed FICT assay is able to detect zoonotic influenza A viruses with a high sensitivity, and it enables the quantitation of the infection intensity by providing the numerical diagnostic values; thus demonstrating enhanced detectability of influenza A viruses.
authors.affiliation: 1. Zoonosis Research Center, Department of Infection Biology, School of Medicine, Wonkwang University, Iksan, Jeonbuk, Republic of Korea.
authors.forename: Seon-Ju
authors.initials: SJ
authors.lastname: Yeo
authors.affiliation: 1. Zoonosis Research Center, Department of Infection Biology, School of Medicine, Wonkwang University, Iksan, Jeonbuk, Republic of Korea.
authors.forename: Dinh Thi
authors.initials: DT
authors.lastname: Huong
authors.affiliation: 1. Zoonosis Research Center, Department of Infection Biology, School of Medicine, Wonkwang University, Iksan, Jeonbuk, Republic of Korea.
authors.forename: Nguyen Ngoc
authors.initials: NN
authors.lastname: Hong
authors.affiliation: 1. Zoonosis Research Center, Department of Infection Biology, School of Medicine, Wonkwang University, Iksan, Jeonbuk, Republic of Korea.
authors.forename: Chun-Ying
authors.initials: CY
authors.lastname: Li
authors.affiliation: 2. Department of Electrical Engineering, Korea Advanced Institute of Science and Technology, Daejeon, Chungnam, Republic of Korea.
authors.forename: Kyunghan
authors.initials: K
authors.lastname: Choi
authors.affiliation: 2. Department of Electrical Engineering, Korea Advanced Institute of Science and Technology, Daejeon, Chungnam, Republic of Korea.
authors.forename: Kyoungsik
authors.initials: K
authors.lastname: Yu
authors.affiliation: 3. Department of Pediatrics, School of Medicine, Wonkwang University, Iksan, Jeonbuk, Republic of Korea.
authors.forename: Du-Young
authors.initials: DY
authors.lastname: Choi
authors.affiliation: 4. GenBody Inc, No.206, Biotech Business IC, DanKook University, San-29, Anseo-dong, Dongnam-gu, Cheonan, Chungnam, Republic of Korea.
authors.forename: Chom-Kyu
authors.initials: CK
authors.lastname: Chong
authors.affiliation: 5. Division of Hematology/Oncology, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, USA.
authors.forename: Hak Soo
authors.initials: HS
authors.lastname: Choi
authors.affiliation: 6. College of Pharmacy, Institute of Pharmaceutical Research and Development, Wonkwang University, Iksan, Jeonbuk, Republic of Korea.
authors.forename: Shyam Kumar
authors.initials: SK
authors.lastname: Mallik
authors.affiliation: 6. College of Pharmacy, Institute of Pharmaceutical Research and Development, Wonkwang University, Iksan, Jeonbuk, Republic of Korea.
authors.forename: Hak Sung
authors.initials: HS
authors.lastname: Kim
authors.affiliation: 7. College of Veterinary Medicine, Kangwon National University, Chunchon, Kangwon, Republic of Korea.
authors.forename: Haan Woo
authors.initials: HW
authors.lastname: Sung
authors.affiliation: 1. Zoonosis Research Center, Department of Infection Biology, School of Medicine, Wonkwang University, Iksan, Jeonbuk, Republic of Korea.
authors.forename: Hyun
authors.initials: H
authors.lastname: Park
mesh_terms: D000818:Animals; D001717:Birds; D002846:Chromatography, Affinity / Q000295:instrumentation / Q000379:methods*; D003374:Coumarins / Q000737:chemistry*; D050091:Dendrimers / Q000737:chemistry*; D005453:Fluorescence; D006801:Humans; D009980:Influenza A virus / Q000737:chemistry / Q000302:isolation & purification*; D005585:Influenza in Birds / Q000175:diagnosis* / Q000821:virology; D007251:Influenza, Human / Q000175:diagnosis* / Q000821:virology; D009698:Nucleoproteins / Q000032:analysis*; D015047:Zoonoses / Q000175:diagnosis* / Q000821:virology
chemical_list: D003374:Coumarins; D050091:Dendrimers; D009698:Nucleoproteins; C030123:coumarin
keywords: Avian influenza A subtype; Conjugation; Coumarin-derived dendrimer; Dot blot immunoassay.; Fluorescent immunochromatographic strip test (FICT); Latex