title: Development and evaluation of a truncated recombinant NS3 antigen-based indirect ELISA for detection of pestivirus antibodies in sheep and goats.
abstract: The aim of this study was to develop an indirect ELISA using the helicase domain of bovine viral diarrhoea virus (BVDV) NS3 protein instead of full-length NS3 protein for detection of BVDV and BDV antibodies in sheep and goats and its validation by comparing its sensitivity and specificity with virus neutralization test (VNT) as the reference test. The purified 50 kDa recombinant NS3 protein was used as the coating antigen in the ELISA. The optimal concentration of antigen was 320 ng/well at a serum dilution of 1:20 and the optimal positive cut-off optical density value was 0.40 based on test results of 418 VNT negative sheep and goat sera samples. When 569 serum samples from sheep (463) and goats (106) were tested, the ELISA showed a sensitivity of 91.71% and specificity of 94.59% with BVDV VNT. A good correlation (93.67%) was observed between the two tests. It showed a sensitivity of 85% and specificity of 86.6% with VNT in detecting BDV antibody positive or negative samples. This study demonstrates the efficacy of truncated recombinant NS3 antigen based ELISA for seroepidemiological study of pestivirus infection in sheep and goats.
authors.affiliation: a High Security Animal Disease Laboratory, Indian Veterinary Research Institute, Anand Nagar , Bhopal , Madhya Pradesh , India.
authors.forename: Semmannan
authors.initials: S
authors.lastname: Kalaiyarasu
authors.forename: Niranjan
authors.initials: N
authors.lastname: Mishra
authors.forename: Katherukamem
authors.initials: K
authors.lastname: Rajukumar
authors.forename: Ram Kumar
authors.initials: RK
authors.lastname: Nema
authors.forename: Sthita Pragnya
authors.initials: SP
authors.lastname: Behera
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood* / Q000276:immunology; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay; D015511:Goat Diseases / Q000097:blood* / Q000276:immunology / Q000821:virology; D006041:Goats; D010576:Pestivirus; D018182:Pestivirus Infections / Q000097:blood* / Q000276:immunology / Q000662:veterinary; D020365:RNA Helicases / Q000737:chemistry / Q000276:immunology; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology; D012697:Serine Endopeptidases / Q000737:chemistry / Q000276:immunology; D012756:Sheep; D012757:Sheep Diseases / Q000097:blood* / Q000276:immunology / Q000821:virology; D017361:Viral Nonstructural Proteins / Q000737:chemistry* / Q000276:immunology; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
chemical_list: D000914:Antibodies, Viral; D020365:RNA Helicases; D011994:Recombinant Proteins; D012697:Serine Endopeptidases; D017361:Viral Nonstructural Proteins; C060028:NS3 protein, flavivirus; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
keywords: BVDV antibodies; NS3 ELISA; VNT; goat; sheep