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
Summary
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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