Identification of T-helper and linear B epitope in the hypervariable region of nucleocapsid protein of PPRV and its use in the development of specific antibodies to detect viral antigen.
Peste des petits ruminants is a highly contagious viral disease of small ruminants making its diagnosis difficult from the similar symptoms of Rinderpest. Computer based prediction algorithms was applied to identify antigenic determinants on the nucleocapsid (N) protein of PPRV. Specificity and antigenicity of each peptide was evaluated by solid phase ELISA. Six specific peptide sequences were evaluated in multiple antigenic peptide (MAP) form and immune response was evaluated by supplementing universal T-helper epitope human IL-1beta peptide (VQGEESNDK, amino acids 163-171). Out of the six peptides 19mer sequence corresponding to 454-472 region of N protein of PPRV was found to be highly immunogenic and specific to PPRV. Evaluation of overlapping peptides differing in length for this 452-472 region, showed minimum length of 14 amino acid residues were required for the stable affinity binding of antigen-antibody. The results of immunization and indirect ELISA indicated the presence of T-helper epitope at the N-terminal end and linear B epitope at the C-terminal region of 454-472 19mer of nucleocapsid peptide of PPRV-nucleocapsid protein. The antipeptide antibodies developed against this region showed specificity to PPRV antigen differentiating it from RPV when used in indirect ELISA and western blot analysis.
Dechamma
H J
HJ
Indian Veterinary Research Institute, Molecular Virology Lab, Bangalore Campus, Hebbal, Bangalore, India. dechammahj@yahoo.com
Dighe
V
V
Kumar
C Ashok
CA
Singh
R P
RP
Jagadish
M
M
Kumar
Satish
S
D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D000956:Antigens, Viral / Q000276:immunology; D015153:Blotting, Western / Q000379:methods / Q000662:veterinary; D022801:Complementarity Determining Regions; D003937:Diagnosis, Differential; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary; D018604:Epitope Mapping; D000939:Epitopes; D018985:Epitopes, B-Lymphocyte / Q000276:immunology*; D018984:Epitopes, T-Lymphocyte / Q000276:immunology*; D005260:Female; D053583:Interleukin-1beta; D019590:Nucleocapsid Proteins / Q000276:immunology*; D029021:Peste-des-Petits-Ruminants / Q000175:diagnosis*; D018107:Peste-des-petits-ruminants virus / Q000276:immunology*; D011817:Rabbits; D011897:Random Allocation; D012301:Rinderpest / Q000175:diagnosis; D012302:Rinderpest virus / Q000276:immunology; D012680:Sensitivity and Specificity
D000914:Antibodies, Viral; D000956:Antigens, Viral; D022801:Complementarity Determining Regions; D000939:Epitopes; D018985:Epitopes, B-Lymphocyte; D018984:Epitopes, T-Lymphocyte; D053583:Interleukin-1beta; D019590:Nucleocapsid Proteins