Theranostics 2014; 4(12):1239-1249. doi:10.7150/thno.10255
Rapid and Quantitative Detection of Zoonotic Influenza A Virus Infection Utilizing Coumarin-derived dendrimer-based Fluorescent Immunochromatographic Strip Test (FICT)
1. Zoonosis Research Center, Department of Infection Biology, School of Medicine, Wonkwang University, Iksan, Jeonbuk, Republic of Korea.
2. Department of Electrical Engineering, Korea Advanced Institute of Science and Technology, Daejeon, Chungnam, Republic of Korea
3. Department of Pediatrics, School of Medicine, Wonkwang University, Iksan, Jeonbuk, Republic of Korea
4. GenBody Inc, No.206, Biotech Business IC, DanKook University, San-29, Anseo-dong, Dongnam-gu, Cheonan, Chungnam, Republic of Korea
5. Division of Hematology/Oncology, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, USA
6. College of Pharmacy, Institute of Pharmaceutical Research and Development, Wonkwang University, Iksan, Jeonbuk, Republic of Korea
7. College of Veterinary Medicine, Kangwon National University, Chunchon, Kangwon, Republic of Korea.
Yeo SJ, Huong DT, Hong NN, Li CY, Choi K, Yu K, Choi DY, Chong CK, Choi HS, Mallik SK, Kim HS, Sung HW, Park H. Rapid and Quantitative Detection of Zoonotic Influenza A Virus Infection Utilizing Coumarin-derived dendrimer-based Fluorescent Immunochromatographic Strip Test (FICT). Theranostics 2014; 4(12):1239-1249. doi:10.7150/thno.10255. Available from http://www.thno.org/v04p1239.htm
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.
Keywords: Avian influenza A subtype, Fluorescent immunochromatographic strip test (FICT), Conjugation, Latex, Coumarin-derived dendrimer, Dot blot immunoassay.