title: Use of hydrophilic extra-viral domain of canine distemper virus H protein for enzyme-linked immunosorbent assay development.
abstract: Simple methods for measuring the levels of serum antibody against canine distemper virus (CDV) would assist in the effective vaccination of dogs. To develop an enzyme-linked immunosorbent assay (ELISA) specific for CDV, we expressed hydrophilic extra-viral domain (HEVD) protein of the A75/17-CDV H gene in a pET 28a plasmid-based Escherichia (E.) coli vector system. Expression was confirmed by dot and Western blotting. We proposed that detection of E. coli-expressed H protein might be conformation- dependent because intensities of the reactions observed with these two methods varied. The H gene HEVD protein was further purified and used as an antigen for an ELISA. Samples from dogs with undetectable to high anti-CDV antibody titers were analyzed using this HEVD-specific ELISA and a commercial CDV antibody detection kit (ImmunoComb). Levels of HEVD antigenicity measured with the assays and immunochromatography correlated. These data indicated that the HEDV protein may be used as antigen to develop techniques for detecting antibodies against CDV.
authors.affiliation: Import Risk Assessment Division, Animal and Plant Quarantine Agency, Anyang 430-757, Korea.
authors.forename: Ki-hyun
authors.initials: KH
authors.lastname: Cho
authors.forename: Jeongmi
authors.initials: J
authors.lastname: Kim
authors.forename: Hyun-ah
authors.initials: HA
authors.lastname: Yoo
authors.forename: Dae-hee
authors.initials: DH
authors.lastname: Kim
authors.forename: Seung-yong
authors.initials: SY
authors.lastname: Park
authors.forename: Chang-seon
authors.initials: CS
authors.lastname: Song
authors.forename: In-soo
authors.initials: IS
authors.lastname: Choi
authors.forename: Joong-bok
authors.initials: JB
authors.lastname: Lee
mesh_terms: D000818:Animals; D000956:Antigens, Viral; D004216:Distemper / Q000175:diagnosis / Q000821:virology*; D004217:Distemper Virus, Canine / Q000276:immunology*; D004283:Dog Diseases / Q000175:diagnosis* / Q000821:virology; D004285:Dogs; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary*; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D006389:Hemagglutinins, Viral
chemical_list: D000956:Antigens, Viral; D006389:Hemagglutinins, Viral
keywords: H gene; canine distemper virus; enzyme-linked immunosorbent assay; hydrophilic extra-viral domain; immunochromatography