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.

Clinical evaluation of rapid fluorescent diagnostic immunochromatographic test for influenza A virus (H1N1)

“…Influenza A/H1N1/2009/CA was provided by Korea Centers for Disease Control and Prevention. It was grown and titrated in eggs as previously described 18 . Influenza A/H3N2 isolated from human in 2014 was donated from Korean National Research Resource Center (Seoul, Korea) (registration number: KBPV-VR-85).…”

Development of a smartphone-based rapid dual fluorescent diagnostic system for the simultaneous detection of influenza A and H5 subtype in avian influenza A-infected patients

“…Avian influenza virus H5N3, H7N1, H1N1, and influenza B were obtained from the Centers for Disease Control & Prevention, Korea. All viruses were cultured by egg inoculation and titrated by a hemagglutination assay as described previously [12]. Next, 400 µL succinic anhydride (60 mg/mL) in 0.1 M sodium phosphate was added to completely exchange -NH2 groups to -COOH groups on the surface of the latex-QD complex which was activated by adding 1.3 µL 50 mM EDC and 10 µL 0.5 M Sulfo-NHS in 0.05 M MES buffer (pH 6.1).…”

“…Given the demand for multiplex testing and the major advantages of the lateral flow assay over alternative rapid diagnostic platforms, this study aimed to adapt the simplicity of the membrane-based lateral flow assay and multiplex techniques. Various fluorescent materials have been applied for lateral flow assays as alternative to conventional gold nanoparticles to detect influenza A as POCT [11][12][13][14][15][16]. Also, self-contained and disposable diagnostic devices have been developed [15,[17][18][19].…”

Advanced nanotechnologies in avian influenza: Current status and future trends – A review

“…This assay provides the benefit of the easy discrimination of the fluorescent emission from the latex beads along with the simple insertion of a long-pass optical filter at a wavelength distinct from the excitation wavelength [92]. In this study, the coumarinderived dendrimers were conjugated with latex beads to obtain fluorescent emissions covering broad output spectral ranges.…”