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Avian Pathol. 2019 Jun 24:1-42. doi: 10.1080/03079457.2019.1634791. [Epub ahead of print]
Novel protein microarray for the detection of avian influenza virus antibodies and simultaneous distinction of antibodies against H5 and H7 subtypes.
Xiao Q1, Bi Z1, Yao L1, Lei J1, Yan Y2, Zhou J2, Yan L1.
Author information
Abstract
Avian influenza virus (AIV) can cause serious zoonotic disease, thereby threatening the poultry industry and human health. An efficient and rapid detection approach is crucial to prevent and control the spread of avian influenza. In this study, a novel protein microarray was developed. Hemagglutininproteins of H5 and H7 subtypes and nucleoprotein (NP) protein were purified and spotted onto the initiator-integrated poly (dimethylsiloxane) as antigens. Monoclonal antibodies with inhibition effect were screened and utilised for the synchronous detection of three avian influenza antibodies in different species. In the protein microarray, the cut-off values were 40%, 50% and 30% inhibition for H5 antibody detection; 50%, 50% and 20% for NP antibody detection; 40%, 50% and 40% for H7 antibody detection in chicken, peacock and duck sera, respectively. The 95 serum samples were detected by microarray, and results were compared with the findings of AIV antibody test enzyme-linked immunosorbent assay (ELISA) or hemagglutination inhibition (HI) test. NP antibody detection in the microarray showed 100% (55/55) agreement ratio in chicken with ELISA. Compared with HI, H5 antibody detection in the microarray showed 100% (95/95) agreement ratio in chicken, peacock and duck, whilst those of H7 displayed 98.18% (54/55) in chicken, 100% (20/20) in peacock and 90% (18/20) in duck. In conclusion, the protein microarray that is first developed is a high-throughput and specific method for the detection of AIV antibodies and simultaneous distinction of antibodies against H5 and H7 subtypes, it can be applied to the serological diagnosis and epidemiological investigation of AIV. Research Highlights A novel protein microarray method has been developed. The microarray can detect AIV antibodies and distinguish between H5 and H7 subtypes. The study lays the foundation for simultaneous identification of multiple pathogens.
KEYWORDS:
AIV; Antibody detection; H5; H7; NP; Protein microarray
PMID: 31232095 DOI: 10.1080/03079457.2019.1634791
Novel protein microarray for the detection of avian influenza virus antibodies and simultaneous distinction of antibodies against H5 and H7 subtypes.
Xiao Q1, Bi Z1, Yao L1, Lei J1, Yan Y2, Zhou J2, Yan L1.
Author information
Abstract
Avian influenza virus (AIV) can cause serious zoonotic disease, thereby threatening the poultry industry and human health. An efficient and rapid detection approach is crucial to prevent and control the spread of avian influenza. In this study, a novel protein microarray was developed. Hemagglutininproteins of H5 and H7 subtypes and nucleoprotein (NP) protein were purified and spotted onto the initiator-integrated poly (dimethylsiloxane) as antigens. Monoclonal antibodies with inhibition effect were screened and utilised for the synchronous detection of three avian influenza antibodies in different species. In the protein microarray, the cut-off values were 40%, 50% and 30% inhibition for H5 antibody detection; 50%, 50% and 20% for NP antibody detection; 40%, 50% and 40% for H7 antibody detection in chicken, peacock and duck sera, respectively. The 95 serum samples were detected by microarray, and results were compared with the findings of AIV antibody test enzyme-linked immunosorbent assay (ELISA) or hemagglutination inhibition (HI) test. NP antibody detection in the microarray showed 100% (55/55) agreement ratio in chicken with ELISA. Compared with HI, H5 antibody detection in the microarray showed 100% (95/95) agreement ratio in chicken, peacock and duck, whilst those of H7 displayed 98.18% (54/55) in chicken, 100% (20/20) in peacock and 90% (18/20) in duck. In conclusion, the protein microarray that is first developed is a high-throughput and specific method for the detection of AIV antibodies and simultaneous distinction of antibodies against H5 and H7 subtypes, it can be applied to the serological diagnosis and epidemiological investigation of AIV. Research Highlights A novel protein microarray method has been developed. The microarray can detect AIV antibodies and distinguish between H5 and H7 subtypes. The study lays the foundation for simultaneous identification of multiple pathogens.
KEYWORDS:
AIV; Antibody detection; H5; H7; NP; Protein microarray
PMID: 31232095 DOI: 10.1080/03079457.2019.1634791