Application of an immunoglobulin Y-alkaline phosphatase bioconjugate as a diagnostic tool for influenza A virus

Bioengineered. 2019 Dec;10(1):33-42. doi: 10.1080/21655979.2019.1586054.

Abstract

The diagnosis of influenza A virus is essential since it can be confused with influenza A like illness and lead to inaccurate drug prescription. In this study, the M2e peptide, a strategic antigen that is conserved in all virus subtypes, was used as a diagnostic marker of influenza A. For the first time, M2e-specific IgY antibody was covalently conjugated to alkaline phosphatase (ALP) enzyme in the presence of glutaraldehyde. The antibody-enzyme bioconjugate was characterized by fluorescence and Fourier-transform infrared spectroscopy. Subsequently, the diagnostic value of this bioconjugate was evaluated by direct sandwich ELISA using nasopharyngeal swab samples positive/negative for H1N1 and H3N2, which were previously analyzed by rRT-PCR for influenza. In conclusion, the M2e-specific IgY-ALP bioconjugate demonstrated positive results for Influenza A in samples that were diagnosed as Influenza A via the RT-PCR method.

Keywords: ELISA methods; Immunoglobulin Y; alkaline phosphatase; diagnosis; influenza A.

MeSH terms

  • Alkaline Phosphatase / chemistry*
  • Alkaline Phosphatase / immunology
  • Amino Acid Sequence
  • Animals
  • Antibodies, Viral / biosynthesis
  • Antibodies, Viral / chemistry*
  • Antibodies, Viral / isolation & purification
  • Antigens, Viral / administration & dosage
  • Antigens, Viral / chemistry
  • Antigens, Viral / immunology*
  • Chickens
  • Cross-Linking Reagents / chemistry
  • Enzyme-Linked Immunosorbent Assay / methods
  • Epitopes / chemistry
  • Female
  • Glutaral / chemistry
  • Humans
  • Immunization
  • Immunoconjugates / chemistry
  • Immunoglobulins / biosynthesis
  • Immunoglobulins / chemistry*
  • Immunoglobulins / isolation & purification
  • Influenza A Virus, H1N1 Subtype / chemistry
  • Influenza A Virus, H1N1 Subtype / immunology
  • Influenza A Virus, H1N1 Subtype / isolation & purification*
  • Influenza A Virus, H3N2 Subtype / chemistry
  • Influenza A Virus, H3N2 Subtype / immunology
  • Influenza A Virus, H3N2 Subtype / isolation & purification*
  • Influenza, Human / diagnosis*
  • Influenza, Human / immunology
  • Influenza, Human / virology
  • Nasopharynx / virology
  • Peptides / administration & dosage
  • Peptides / chemistry
  • Peptides / immunology
  • Spectroscopy, Fourier Transform Infrared

Substances

  • Antibodies, Viral
  • Antigens, Viral
  • Cross-Linking Reagents
  • Epitopes
  • IgY
  • Immunoconjugates
  • Immunoglobulins
  • Peptides
  • Alkaline Phosphatase
  • Glutaral

Grants and funding

This study was supported by the Scientific and Technological Research Council of Turkey with the project titled ‘The development of a rapid diagnostic kit using the immunochromatographic method of depending on an M2e peptide-specific IgY antibody for the diagnosis of influenza A infection’ (Project No: 115S132); The Scientific and Technological Research Council of Turkey [115S132].