4.7 Article

Development of a Rapid Fluorescent Diagnostic System to Detect Subtype H9 Influenza A Virus in Chicken Feces

Journal

Publisher

MDPI
DOI: 10.3390/ijms22168823

Keywords

fecal specimen; subtype H9 influenza A virus; rapid fluorescent immunochromatographic test; monoclonal antibody

Funding

  1. Bio & Medical Technology Development Program of the National Research Foundation (NRF) - Korean government (MSIT) [2018M3A9H4055767, NRF-2018M3A9H4055194]
  2. Priority Research Centers Program through the National Research Foundation of Korea (NRF) - Ministry of Education [NRF-2015R1A6A1A03032236]
  3. Basic Science Research Program through the National Research Foundation of Korea(NRF) - Ministry of Education [2021R1I1A1A01049073]
  4. National Research Foundation of Korea [2021R1I1A1A01049073, 2018M3A9H4055194] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)

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Novel monoclonal antibodies (McAbs) against influenza A H9 viruses were developed for the rapid diagnostic detection of H9N2 virus in feces samples, with linear epitope McAbs showing superior performance in animal studies with 100% relative sensitivity in fecal samples at 6 days post-infection.
The circulation of the H9N2 virus results in significant economic losses in the poultry industry, and its zoonotic transmission highlights the need for a highly sensitive and rapid diagnostic and detection system for this virus. In this study, the performance of lateral flow test strips for a fluorescent immunochromatographic test (FICT) was optimized for the diagnosis of H9N2 virus-infected animal samples. The novel monoclonal antibodies (McAbs) against influenza A H9 viruses were developed, and two categories of McAbs with linear and conformational epitopes were compared for the performance of rapid diagnostic performance in the presence of feces sample at different time points (2, 4, and 6 days) post-infection (dpi). The limit of detection (LOD) of FICT and Kd values were comparable between linear and conformational epitope McAbs. However, superior performance of linear epitope McAbs pairs were confirmed by two animal studies, showing the better diagnostic performance showing 100% relative sensitivity in fecal samples at 6 dpi although it showed less than 80% sensitivity in early infection. Our results imply that the comparable performance of the linear epitope McAbs can potentially improve the diagnostic performance of FICT for H9N2 detection in feces samples. This highly sensitive rapid diagnostic method can be utilized in field studies of broiler poultry and wild birds.

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