Reliable Luciferase Immunoprecipitation System Assays for the Detection of Porcine Reproductive and Respiratory Syndrome Virus Antibodies.

LIPS assays detect PRRSV antibodies earlier and more reliably than commercial ELISA.

This study presents the development and validation of two luciferase immunoprecipitation system (LIPS) assays for detecting porcine reproductive and respiratory syndrome virus (PRRSV) antibodies, targeting the nsp7 protein. The assays utilize Gaussia luciferase (GLuc-nsp7) and Nano luciferase with N-terminal signal peptide (GNLuc-nsp7) as reporter proteins. Optimal performance was achieved with 10⁷ luminance units of fusion antigen and 1:50 serum dilution, yielding high positive-to-negative ratios. Both assays demonstrated excellent specificity with no cross-reactivity against sera from pigs infected with other common pathogens. A significant advancement over commercial methods was observed, with LIPS assays detecting PRRSV-specific antibodies as early as 3 days post-challenge, compared to 8 days for the IDEXX ELISA. The GNLuc-nsp7 variant shows particular promise for on-farm applications due to its simplified protocol eliminating automated substrate requirements. These LIPS assays represent reliable diagnostic tools suitable for both laboratory and field implementations, potentially improving early detection and biosecurity measures in swine production systems.

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