title: Detection of circulating Asian H5N1 viruses by a newly established monoclonal antibody.
abstract: Monoclonal antibodies (MAbs) against the recently emerged Asian H5N1 virus (A/crow/Kyoto/53/2004) were generated. From five anti-hemagglutinin (HA) MAbs, four antibodies (3C11, 4C12, 3H12, and 3H4) broadly in vitro recognized and neutralized H5 subtypes, including H5N1. By contrast, the 4G6 MAb specifically reacted with H5N1-HA and not with H5N2- or H5N3-HAs from previous epidemics. The 4G6 MAb was useful for immunofluorescence assays but not for immunoblotting, suggesting that this antibody recognizes a conformational epitope of the H5N1-HA protein. An intensive epitope-mapping analysis demonstrated that the 4G6 MAb recognizes Asp59, which is highly conserved among currently circulating H5N1 lineages. Further, a 4G6-based antigen capture enzyme-linked immunosorbent assay detected H5N1 even that derived from clade 2.2 (A/chicken/Egypt/CL-61/2007) from infected chicken lung before virus isolation. Taken together, these results suggest that the established MAbs, especially 4G6, are useful for rapid and specific detection of Asian H5N1 viruses.
authors.affiliation: International Research Center for Infectious Diseases, Research Institute for Microbial Diseases, Osaka University, Suita, Osaka 565-0871, Japan.
authors.forename: Anariwa
authors.initials: A
authors.lastname: Du
authors.forename: Tomo
authors.initials: T
authors.lastname: Daidoji
authors.forename: Takaaki
authors.initials: T
authors.lastname: Koma
authors.forename: Madiha S
authors.initials: MS
authors.lastname: Ibrahim
authors.forename: Shota
authors.initials: S
authors.lastname: Nakamura
authors.forename: U Chandimal
authors.initials: UC
authors.lastname: de Silva
authors.forename: Mayo
authors.initials: M
authors.lastname: Ueda
authors.forename: Cheng-Song
authors.initials: CS
authors.lastname: Yang
authors.forename: Teruo
authors.initials: T
authors.lastname: Yasunaga
authors.forename: Kazuyoshi
authors.initials: K
authors.lastname: Ikuta
authors.forename: Takaaki
authors.initials: T
authors.lastname: Nakaya
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology*; D001208:Asia; D002645:Chickens / Q000821:virology; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D005260:Female; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000145:classification / Q000276:immunology*; D053124:Influenza A Virus, H5N1 Subtype / Q000276:immunology / Q000302:isolation & purification*; D005585:Influenza in Birds / Q000175:diagnosis* / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D009500:Neutralization Tests; D010802:Phylogeny
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D019267:Hemagglutinin Glycoproteins, Influenza Virus; C109963:hemagglutinin, avian influenza A virus