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Development of a colloidal gold-based immunochromatographic strip test using two monoclonal antibodies to detect H7N9 avian influenza virus

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Abstract

H7N9 low pathogenic avian influenza viruses (AIVs) emerged in China in 2013 and mutated into highly pathogenic strains in 2017, causing disease in infected birds and humans. Thus, the development of rapid, specific, and sensitive detection methods is urgently required. Herein, two specific monoclonal antibodies against H7N9 AIV were produced to develop a colloidal gold-based immunochromatographic test strip to detect H7N9 AIV. High specificity, repeatability, and sensitivity were achieved, with a detection limit of two hemagglutinin units or 102.55 50% tissue culture infective dose. This assay may represent a powerful tool to rapidly detect H7N9 influenza viruses in the future.

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Funding

This study was funded by Grants from the National Science Foundation of the People’s Republic of China (81502852), the National Science and Technology Major Project for the Control and Prevention of Major Infectious Diseases in China (2018ZX10711001, 2018ZX10102001 and 2020ZX09001016-004-002), Zhejiang Provincial Natural Science Foundation of China (LY19H260006), and the Independent Task of State Key Laboratory for Diagnosis and Treatment of Infectious Diseases (2019ZZ17).

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Authors and Affiliations

  1. State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, the First Affiliated Hospital, School of Medicine, Zhejiang University, 79 Qingchun Road, Hangzhou, 310003, China

    Fan Yang, Yixin Xiao, Bin Chen, Liyan Wang, Fumin Liu, Hangping Yao, Nanping Wu & Haibo Wu

  2. National Clinical Research Center for Infectious Diseases, the First Affiliated Hospital, School of Medicine, Zhejiang University, 79 Qingchun Road, Hangzhou, 310003, China

    Fan Yang, Yixin Xiao, Bin Chen, Liyan Wang, Fumin Liu, Hangping Yao, Nanping Wu & Haibo Wu

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  1. Fan Yang
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  2. Yixin Xiao
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  3. Bin Chen
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Correspondence to Haibo Wu.

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The authors declare that they have no conflict of interest.

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The animal experiment was approved by the Institutional Animal Care Use Committee (IACUC) of Zhejiang University (No. 2017-015).

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Edited by Takeshi Noda.

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Supplementary file1 (DOCX 14 kb)

11262_2020_1742_MOESM2_ESM.tif

Fig S1. A schematic diagram of the immunochromatographic strip. The strip consists of three regions: 1) a sample placement region; 2) a reagent pad region; and 3) a result region. The reagent pad contains Platinum-gold-colloid conjugated MAb 2D5. The result region contains nitrocellulose membrane-immobilized mAb 2F5 and anti-mouse polyclonal antibodies, which serve as the capture antibodies in the H7-positive and control lines, respectively. (TIF 503 kb)

11262_2020_1742_MOESM3_ESM.png

Fig S2. The location of escape residue displayed on the crystal structure of the H7 trimer from A/Shanghai/02/2013 (PDB: 4NL6). The two amino acid residues recognized by the mAb 2D5 (G131E) and 2F5 (E124L) are highlighted in red and purple, respectively. Compared the HA gene of A/chicken/Zhejiang/DTID-ZJU01/2013(H7N9) with the A/Shanghai/02/2013(H7N9), the sequences of these two viruses are highly similar (>99.99%). The epitopes of 2D5 and 2F5 were showed on a space-filling model of the trimeric membrane-distal globular head of A/Shanghai/02/2013(H7N9). (PNG 185 kb)

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Yang, F., Xiao, Y., Chen, B. et al. Development of a colloidal gold-based immunochromatographic strip test using two monoclonal antibodies to detect H7N9 avian influenza virus. Virus Genes 56, 396–400 (2020). https://doi.org/10.1007/s11262-020-01742-8

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