Abstract

Matrix M protein combined with nucleocapsid N protein could be a promising combination of virus antigens for diagnosing the porcine reproductive and respiratory syndrome. The goal of this work was to express the recombinant M protein of the porcine reproductive and respiratory syndrome virus inEscherichia colicells and compare its serological reactivity with the N protein of the virus. The gene coding for the M protein was cloned into the pDest17 vector. The resulting protein was purified by metalochelating affinity chromatography. Recombinant M protein was applied as an antigen in immunoblot test and compared on a panel of porcine sera with N protein based IDEXX test. Of 120 examined samples, the majority (78.3%) gave identical results using both compared tests. From the group of discrepant results, IDEXX test identified considerably more positive sera (17.5%) than M protein based test (4.2%). The main contribution of the work is finding that although IDEXX test proved to be more sensitive than M protein based test, 4.2% of sera would escape detection by serological test based on N protein. Further development and purification of the M protein for the use in Enzyme Linked Immunosorbent Assay format test could increase the performance of serological testing.

Highlights

  • Matrix M protein combined with nucleocapsid N protein could be a promising combination of virus antigens for diagnosing the porcine reproductive and respiratory syndrome

  • The 522 bp long fragment of open reading frames (ORFs) 6 gene was RT-polymerase chain reaction (PCR) amplified from genomic ribonucleic acid (RNA) of PRRS virus strain and successfully cloned into pENTRTM Vector by TOPO cloning

  • The PRRS virus nucleoprotein alone is considered as a reliable antigen, in some cases nonspecific reactions can be seen and enrichment of serological testing by additional antigens could be beneficial

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Summary

Introduction

Matrix M protein combined with nucleocapsid N protein could be a promising combination of virus antigens for diagnosing the porcine reproductive and respiratory syndrome. The goal of this work was to express the recombinant M protein of the porcine reproductive and respiratory syndrome virus in Escherichia coli cells and compare its serological reactivity with the N protein of the virus. Recombinant M protein was applied as an antigen in immunoblot test and compared on a panel of porcine sera with N protein based IDEXX test. From the group of discrepant results, IDEXX test identified considerably more positive sera (17.5%) than M protein based test (4.2%). The porcine reproductive and respiratory syndrome virus (PRRSV) causes one of the most threatening diseases for pig industry world-wide.

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