The overall purpose of this project is to institute a collaboration between Kansas State University (KSU), South Dakota State University (SDSU), and Iowa State University (ISU) to develop and implement a new serological tool, known as Luminex, for the simultaneous detection of PRRSV, PCV2 and SIV antibodies in serum and oral fluid samples. This technology represents a “faster, better and cheaper” alternative to traditional serological tests and can be extended to include the analysis of up to 50 antigens or pathogens in a single sample. Previous projects funded by NPB have demonstrated a proof of concept in the capacity of Luminex to detect antibodies. The goal of this project was to transfer this technology into a workable assay kit for standardization across diagnostic labs. The principal objective was the establishment of samples and standard methodologies for PRRSV, SIV and PCV2 Luminex assays. Many samples were already available in the PI and co-PI labs. However, one specific objective was the creation of a set of samples from pigs infected with a European-like type 1 PRRS virus. As part of the development, the investigators engaged a private biologics company to prepare a commercial kit, which is based on the technology and reagents provided by the project investigators. A commercial test is the best means to establish a standardized test for use by all veterinary diagnostic labs. Recently, USDA recognized the collaborative approach towards the development of a commercial assay as having a high impact.
Objective 1. Establish a panel of serum and oral fluid samples for the standardization of PRRSV, SIV and PCV2 Luminex assays. The purpose of this objective was to create a panel of samples with known antibody reactivity. Standards would include oral fluid and serum for detection of IgM, IgG, and IgA isotypes. Funding was specifically requested to develop a sample set from PRRSV type I genotype-infected pigs.
Objective 2. Determine test reproducibility. The purpose of this objective was to assay the panel of standard samples in Objective 1 and compare results across laboratories (reproducibility).
Objective 3. Conduct a large scale inter-laboratory field validation of PRRSV N-based Luminex assay for the detection of IgM and IgG in serum, and IgM, IgG and IgA in oral fluid samples. The purpose of this objective was to conduct a field validation of PRRSV N-based Luminex assays using serum and oral fluid samples from herds of known PRRSV serological status.