Novel application of Influenza A virus-inoculated chorioallantoic membrane to characterize a NP-specific monoclonal antibody for immunohistochemistry assaying
Journal of Veterinary Science
;
: 51-57, 2019.
Artigo
em Inglês
| WPRIM
| ID: wpr-758884
ABSTRACT
Monoclonal antibodies (MAbs) are widely applied in disease diagnoses. Herein, we report a MAb, WF-4, against Influenza A virus nucleoprotein (NP), its broad response with Influenza A virus, and its application in an immunohistochemistry (IHC) assay. WF-4 was screened by immunofluorescence assay (IFA). The results showed that its reactivity with baculovirus-expressed full-length recombinant NP (rNP) in Western blot (WB), indicating its IHC applicability. Fifteen Influenza A virus (reference subtypes H1 to H15) infected chicken embryonated chorioallantoic membranes (CAM), fixed by formalin, were all detectable in the WF-4-based IHC assay. Also, the reactivity of the IHC test with NP from experimentally inoculated H6N1 and from all recent outbreaks of H5 subtype avian Influenza A virus (AIV) field cases in Taiwan showed positive results. Our data indicate that CAM, a by-product of Influenza A virus preparation, is helpful for Influenza A virus-specific MAb characterization, and that the WF-4 MAb recognizes conserved and linear epitopes of Influenza A virus NP. Therefore, WF-4 is capable of detecting NP antigens via IHC and may be suitable for developing various tests for diagnosis of Influenza A virus and, especially, AIV infection.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Vírus da Influenza A
/
Taiwan
/
Imuno-Histoquímica
/
Galinhas
/
Western Blotting
/
Surtos de Doenças
/
Imunofluorescência
/
Membrana Corioalantoide
/
Diagnóstico
/
Influenza Humana
Tipo de estudo:
Estudo diagnóstico
Limite:
Animais
País/Região como assunto:
Ásia
Idioma:
Inglês
Revista:
Journal of Veterinary Science
Ano de publicação:
2019
Tipo de documento:
Artigo
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