Use of hydrophilic extra-viral domain of canine distemper virus H protein for enzyme-linked immunosorbent assay development.
10.4142/jvs.2014.15.4.503
- Author:
Ki Hyun CHO
1
;
Jeongmi KIM
;
Hyun Ah YOO
;
Dae Hee KIM
;
Seung Yong PARK
;
Chang Seon SONG
;
In Soo CHOI
;
Joong Bok LEE
Author Information
1. Import Risk Assessment Division, Animal and Plant Quarantine Agency, Anyang 430-757, Korea.
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- Keywords:
canine distemper virus;
enzyme-linked immunosorbent assay;
H gene;
hydrophilic extra-viral domain;
immunochromatography
- MeSH:
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
- From:Journal of Veterinary Science
2014;15(4):503-509
- CountryRepublic of Korea
- Language:English
-
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.
