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KMID : 0893420140150040503
Journal of Veterinary Science
2014 Volume.15 No. 4 p.503 ~ p.509
Use of hydrophilic extra-viral domain of canine distemper virus H protein for enzyme-linked immunosorbent assay development
Cho Ki-Hyun

Kim Jeong-Mi
Yoo Hyun-Ah
Kim Dae-Hee
Park Seung-Yong
Song Chang-Seon
Choi In-Soo
Lee Joong-Bok
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
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