title: Development of a recombinant N-Gp5c fusion protein-based ELISA for detection of antibodies to porcine reproductive and respiratory syndrome virus.
abstract: The recent dramatic increase in reported cases of porcine reproductive and respiratory syndrome (PRRS) in pig farms is a potential threat to the global swine industry, and thus, detecting PRRS virus (PRRSV) in pig herds is essential to help control the spread of PRRS. IDEXX HerdChek™ PRRS, a commercially available indirect enzyme-linked immunosorbent assay (iELISA), is the industry standard for detection of antibodies against PRRSV. In the present study, an effective iELISA for detection of PRRSV antibodies was developed using a recombinant fusion protein N-Gp5c (rN5c, a combination of the nucleocapsid protein and the C-terminal 78 aa of Gp5) produced in Escherichia coli. This assay was validated by comparison with an immunofluorescent assay and IDEXX-ELISA. The diagnostic specificity, sensitivity, and accuracy of the rN5c-iELISA method were 94.8, 95.6, and 95.1%, respectively. Cross-reactivity assays demonstrated that iELISA was PRRSV-specific. Repeatability tests revealed that the coefficients of variation of positive sera within and between runs were less than 13 and 22%, respectively. The rN5c-iELISA is simpler to produce and perform, time-saving, and suitable for large scale surveys of PRRSV infection at lower cost.
authors: Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100101, China.
authors: Caiwei
authors: C
authors: Chen
authors: Wenhui
authors: W
authors: Fan
authors: Xiaojuan
authors: X
authors: Jia
authors: Jing
authors: J
authors: Li
authors: Yuhai
authors: Y
authors: Bi
authors: Wenjun
authors: W
authors: Liu
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood* / Q000276:immunology; D001483:Base Sequence; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D004926:Escherichia coli / Q000378:metabolism; D019590:Nucleocapsid Proteins / Q000276:immunology; D019318:Porcine Reproductive and Respiratory Syndrome / Q000175:diagnosis* / Q000276:immunology* / Q000821:virology; D019316:Porcine respiratory and reproductive syndrome virus / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000276:immunology; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis / Q000821:virology; D014759:Viral Envelope Proteins / Q000276:immunology
chemical_list: D000914:Antibodies, Viral; D019590:Nucleocapsid Proteins; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; C114789:glycoprotein 5, PRRSV