Abstract
BackgroundThe Nucleoprotein (NP) is the most abundant and highly immunogenic protein in canine distemper virus (CDV), playing an important role in CDV viral replication and assembly.ResultsIn this study, a specific monoclonal antibody, named C8, was produced against the NP protein C terminal (amino acids 401–523). A linear N protein epitope was identified by subjecting a series of partially overlapping synthesized peptides to enzyme-linked immunosorbent assay (ELISA) analysis.The results indicated that 444GDKYPIHFNDER455 was the minimal linear epitope that could be recognized by mAb C8. Sequence alignments demonstrated that this linear epitope is less conserved among three CDV genotypes. We next analyzed the level of conservation of the defined epitope in19 Chinese CDV clinical isolates, and it has one site variation in amino acid among these CDV isolations. 2 isolates have the amino acid mutations F451L, while one has P448Ssubstitution.Phylogenetic analysis showed the two isolates with F451Lsubstitution had a closer relationship in a virulent strain ZJ-7, so the epitope may be a significant tag associated with virus virulence.ConclusionThis collection of mAb along with defined linear epitope may provide useful reagents for investigations of NP protein function and the development of CDV specific diagnostics.
Highlights
The Nucleoprotein (NP) is the most abundant and highly immunogenic protein in canine distemper virus (CDV), playing an important role in CDV viral replication and assembly
Canine distemper virus (CDV) is an enveloped negativestrand RNA virus classified into the genus Morbillivirus within the family Paramyxoviridae [1]
All experimental protocols were approved by the Review Board Institute of Special Animal and Plant Sciences, Chinese Academy of Agricultural Sciences (CAAS)
Summary
The Nucleoprotein (NP) is the most abundant and highly immunogenic protein in canine distemper virus (CDV), playing an important role in CDV viral replication and assembly. Nucleoprotein (NP) is the most abundant and highly immunogenic protein in Morbillivirus, and is an internal protein which packs the viral RNA genome and forms a helical nucleocapsid. It can be used as the basis of most diagnostic assays for CDV presently [4]. The interest in diagnostic applications of CDV NP has focused attention on identifying more antigenic epitopes, especially B cell. We decided to perform the specific mAbs and identify their epitopes in CDVNP C-terminal region precisely
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