Development of recombinant nucleocapsid protein based IgM-ELISA for the early detection of distemper infection in dogs.
An IgM-ELISA based on a 16-kDa recombinant protein produced for the conserved and functional middle region of nucleocapsid protein of Canine distemper virus was developed. Out of 70 serum samples from distemper-suspected and vaccinated dogs analyzed, 34 serum samples (49%) were positive. The specificity of this ELISA was confirmed by blocking and adsorption experiments. The IgM-ELISA based on the recombinant nucleocapsid protein showed a strong correlation (r=0.857, p<0.0001 at 95% CI) and good agreement (kappa=0.714) with the conventional Vero cell culture distemper antigen based IgM-ELISA. The percent positivity was more in dogs with systemic signs (62%) by recombinant nucleocapsid protein IgM-ELISA. Out of 70 clinical serum samples, 69 samples were used along with 4 control sera used in the IgM-ELISA for the detection of viral RNA by Slot blot hybridization and 26 of them (36%) were positive. Fifty-one percent agreement was observed between the recombinant nucleocapsid protein IgM-ELISA and Slot blot hybridization. The analysis of clinical history of the dogs with systemic signs supported the application of IgM-ELISA over Slot blot hybridization in the early detection of distemper infection.
Centre for Biotechnology, Anna University, Guindy, Chennai 600025, India.
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Latha
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Geetha
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Ramadass
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Narayanan
D000818:Animals; D000914:Antibodies, Viral; D000956:Antigens, Viral; D004216:Distemper / Q000097:blood / Q000175:diagnosis* / Q000276:immunology*; D004217:Distemper Virus, Canine / Q000276:immunology*; D004285:Dogs; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary*; D015151:Immunoblotting / Q000662:veterinary; D007075:Immunoglobulin M / Q000276:immunology*; D019590:Nucleocapsid Proteins / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology*; D046509:Tissue Culture Techniques
D000914:Antibodies, Viral; D000956:Antigens, Viral; D007075:Immunoglobulin M; D019590:Nucleocapsid Proteins; D011994:Recombinant Proteins