Expression and diagnostic use of recombinant M protein of the porcine reproductive and respiratory syndrome virus
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Published:2017
Issue:1
Volume:86
Page:11-17
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ISSN:0001-7213
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Container-title:Acta Veterinaria Brno
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language:en
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Short-container-title:Acta Vet. Brno
Author:
Frölichová Jitka,Molinková Dobromila,Sedlinská Markéta,Celer Vladimír
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.
Funder
Ministerstvo Školství, Mládeže a Tělovýchovy
Veterinární a Farmaceutická Univerzita Brno
Národní Agentura pro Zemědělsk Vzkum
Publisher
University of Veterinary and Pharmaceutical Sciences
Subject
General Veterinary