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  • Title: Development of a recombinant N-Gp5c fusion protein-based ELISA for detection of antibodies to porcine reproductive and respiratory syndrome virus.
    Author: Chen C, Fan W, Jia X, Li J, Bi Y, Liu W.
    Journal: J Virol Methods; 2013 Apr; 189(1):213-20. PubMed ID: 23439032.
    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.
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