Expression and diagnostic use of recombinant M protein of the porcine reproductive and respiratory syndrome virus

To chci

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. … celý popis

Podrobná bibliografie

Hlavní autor:

Jitka Frölichová

Další autoři:

Dobromila Molinková, 1973-
Markéta Sedlinská
Vladimír Celer, 1965-

Typ dokumentu:

Články

Publikováno v:

Acta veterinaria. -- ISSN 0001-7213. -- Roč. 86, č. 1 (2017), s. 11-17

Témata:

SWINE

ANIMAL DISEASES

INFECTIOUS DISEASES

VIROSES

PRRS

Diagnostic techniques

serology

ANTIGENS

RECOMBINANT PROTEINS

VIRUSES

GENES

CLONING

CELL CULTURE

ESCHERICHIA COLI

VECTORS

CHROMATOGRAPHY

IMMUNOBLOTTING

ELISA

TESTING

prasata

choroby zvířat

infekční choroby

virové nemoci

diagnostické metody

sérologie

antigeny

rekombinantní bílkoviny

viry

geny

klonování

buněčná kultura

Escherichia coli

vektory

chromatografie

immunoblotting

testování

IDEXX test

Popis jednotky:

1 obrázek, 1 tabulka

Fyzický popis:

Ilustrace

Bibliografie:

Literatura na s. 16-17