title: Development of a monoclonal antibody based competitive-ELISA for detection and titration of antibodies to peste des petits ruminants (PPR) virus.
abstract: Peste des petits ruminants (PPR) is an acute febrile, viral, disease of small ruminants with great economic importance. A competitive-ELISA (c-ELISA) test was developed for detection of antibodies to PPR virus in the sera samples of goats and sheep. The test uses monoclonal antibody to a neutralizing epitope of haemagglutinin protein of the virus. Based on the distribution of known negative sera samples (n=933) in respect of PPR virus antibodies in the test, a cut-off value was set as 38%. This value was the result of mean of negative population added with two times the standard deviations. A total of 1668 sera samples from goat and sheep and 32 sera from cattle were screened by c-ELISA and virus neutralization test (VNT). Efficacy of c-ELISA compared very well with VNT having high relative specificity (98.4%) and sensitivity (92.4%). The sensitivity of c-ELISA for PPR sero-surveillance could further be increased (95.4%), if the target population is non-vaccinated. c-ELISA test correlated well with VNT (r=0.845) for end-point titration of PPR virus antibody in 64 goat sera samples. It could clearly separate infected population from uninfected in field sera. Using c-ELISA test paired sera samples from 13 goats provided a clear diagnosis of PPR virus infection. Furthermore, antibodies to PPR virus could be successfully detected during 1 year after vaccination in four goats inoculated with an experimental PPR vaccine. Findings suggest that the c-ELISA test developed can easily replace VNT for sero-surveillance, sero-monitoring, diagnosis from paired sera samples and end-point titration of PPR virus antibodies.
authors.affiliation: Division of Virology, Indian Veterinary Research Institute, Mukteswar, Nainital 263 138, Uttaranchal, India.
authors.forename: R P
authors.initials: RP
authors.lastname: Singh
authors.forename: B P
authors.initials: BP
authors.lastname: Sreenivasa
authors.forename: P
authors.initials: P
authors.lastname: Dhar
authors.forename: L C
authors.initials: LC
authors.lastname: Shah
authors.forename: S K
authors.initials: SK
authors.lastname: Bandyopadhyay
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral / Q000097:blood; D000956:Antigens, Viral; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D015511:Goat Diseases / Q000097:blood / Q000821:virology*; D006041:Goats; D009500:Neutralization Tests / Q000662:veterinary; D029021:Peste-des-Petits-Ruminants / Q000097:blood / Q000821:virology*; D018107:Peste-des-petits-ruminants virus / Q000276:immunology*; D012680:Sensitivity and Specificity; D012756:Sheep; D012757:Sheep Diseases / Q000097:blood / Q000821:virology*
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral