J Vet Sci.  2014 Dec;15(4):503-509. 10.4142/jvs.2014.15.4.503.

Use of hydrophilic extra-viral domain of canine distemper virus H protein for enzyme-linked immunosorbent assay development

Affiliations
  • 1Import Risk Assessment Division, Animal and Plant Quarantine Agency, Anyang 430-757, Korea.
  • 2ISU Abxis Co., Ltd. Yonsei University Medical Center, Seoul 120-752, Korea.
  • 3Department of Infectious Diseases, College of Veterinary Medicine, Konkuk University, Seoul 143-701, Korea.
  • 4Veterinary Science Research Institute, College of Veterinary Medicine, Konkuk University, Seoul 143-701, Korea. virus@konkuk.ac.kr

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.

Keyword

canine distemper virus; enzyme-linked immunosorbent assay; H gene; hydrophilic extra-viral domain; immunochromatography

MeSH Terms

Animals
Antigens, Viral/*diagnostic use
Distemper/diagnosis/*virology
Distemper Virus, Canine/*immunology
Dog Diseases/*diagnosis/virology
Dogs
Enzyme-Linked Immunosorbent Assay/*veterinary
Escherichia coli/genetics
Genetic Vectors/genetics
Hemagglutinins, Viral/*diagnostic use
Antigens, Viral
Hemagglutinins, Viral
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