Validation efficiency of coupling of beads and probes, used in high-throughput multiplexed nucleic acid detection of respiratory and reproductive pathogens in swine

This file set contains results files from the Bio-Plex integrated biomolecular assay system, each containing the data from a reading, the Protocol parameters used to collect that data and analysis tools for interpreting the data. The .rbx format data files must be opened by the Bio-Plex Manager integrated software. Please see the references link below for .xls format data related to this study.<br><br>Magnetic carboxylic beads were coupled to different virus probes as follows: PRV (no. 26), PRRSV (no. 29), CSFV (no. 34), JEV (no. 36), PCV-2 (no. 46), ASFV (no. 62) and PPV (no. 65). <br>Validation of coupling was performed with different amounts of biotin-modified RCS. The curve was drawn with 0, 5, 10, 20, 50, 100 and 200 fmol RCS, respectively.<br>Article abstract:<br>Background: The aim of this study was to develop a multiple PCR assay based on the suspension array system for the simultaneous detection of respiratory and reproductive pathogens in swine. Pseudorabies virus (PRV), Japanese encephalitis virus (JEV), classic swine fever virus (CFSV), African swine fever virus (ASFV), porcine circovirus type 2 (PCV-2), porcine reproductive and respiratory syndrome virus (PRRSV) and porcine parvovirus (PPV) are the major respiratory and reproductive viral pathogens in pig farms.<br>Results: Seven pairs of specific primers and probes were designed, and multiple PCR was performed, with the PCR products hybridized to beads coupled to probes, which were then detected by Bio-Plex suspension array system. The limit of detection, specificity and repeatability of this method was determined. The assay was further tested using 137 clinical samples, and the results were compared with conventional PCR to evaluate the ability of the method to diagnose porcine viruses. The results showed that the assay had a high degree of specificity and repeatability, and the simultaneous detection limit for the seven viruses reached 103 copies/μL. The detection of viruses in clinical samples using the Bio-Plex method was high and comparable to conventional PCR (&gt; 90%).