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Expression and diagnostic use of recombinant M protein of the porcine reproductive and respiratory syndrome virus
Jitka Frölichová, Dobromila Molinková, Markéta Sedlinská, Vladimír Celer
Jazyk angličtina Země Česko
Typ dokumentu přehledy
Open Access Digital Library od 1978-01-01
ROAD: Directory of Open Access Scholarly Resources od 1978
- MeSH
- Escherichia coli genetika izolace a purifikace metabolismus MeSH
- reprodukční a respirační syndrom prasat * genetika klasifikace MeSH
- sérologie MeSH
- Sus scrofa MeSH
- Publikační typ
- přehledy MeSH
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. 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.
Literatura
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- $a Frölichová, Jitka $7 _AN095013 $u University of Veterinary and Pharmaceutical Sciences Brno, Faculty of Veterinary Medicine, Department of Infectious Diseases and Microbiology, Brno, Czech Republic
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- $a 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. 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.
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