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

被引:1
作者
Frolichova, Jitka [1 ]
Molinkova, Dobromila [1 ,3 ]
Sedlinska, Marketa [2 ]
Celer, Vladimir [1 ,3 ]
机构
[1] Univ Vet & Pharmaceut Sci Brno, Fac Vet Med, Dept Infect Dis & Microbiol, Palackeho Tr 1946-1, Brno 61242, Czech Republic
[2] Univ Vet & Pharmaceut Sci Brno, Fac Vet Med, Clin Horse Dis, Brno, Czech Republic
[3] Univ Vet & Pharmaceut Sci Brno, Fac Vet Med, CEITEC Cent European Inst Technol, Brno, Czech Republic
关键词
PRRS; pigs; matrix protein; expression; E; coli; serology; MAJOR STRUCTURAL PROTEINS; SYNDROME PRRS VIRUS; LELYSTAD-VIRUS; BACULOVIRUS EXPRESSION;
D O I
10.2754/avb201786010011
中图分类号
S85 [动物医学(兽医学)];
学科分类号
0906 ;
摘要
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.
引用
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页码:11 / 17
页数:7
相关论文
共 18 条
[1]  
Cavanagh D, 1997, ARCH VIROL, V142, P629
[2]   MOLECULAR CHARACTERIZATION OF PORCINE REPRODUCTIVE AND RESPIRATORY SYNDROME VIRUS, A MEMBER OF THE ARTERIVIRUS GROUP [J].
CONZELMANN, KK ;
VISSER, N ;
VANWOENSEL, P ;
THIEL, HJ .
VIROLOGY, 1993, 193 (01) :329-339
[3]   Current knowledge on the structural proteins of porcine reproductive and respiratory syndrome (PRRS) virus: comparison of the North American and European isolates [J].
Dea, S ;
Gagnon, CA ;
Mardassi, H ;
Pirzadeh, B ;
Rogan, D .
ARCHIVES OF VIROLOGY, 2000, 145 (04) :659-688
[4]   Baculovirus expression of proteins of porcine reproductive and respiratory syndrome virus strain Olot/91. Involvement of ORF3 and ORF5 proteins in protection [J].
Duran J.P. ;
Climent I. ;
Sarraseca J. ;
Urniza A. ;
Cortés E. ;
Vela C. ;
Casal J.I. .
Virus Genes, 1997, 14 (1) :19-29
[5]   AN OUTBREAK OF BLUE-EARED PIG-DISEASE (PORCINE REPRODUCTIVE AND RESPIRATORY SYNDROME) IN 4 PIG HERDS IN GREAT-BRITAIN [J].
HOPPER, SA ;
WHITE, MEC ;
TWIDDY, N .
VETERINARY RECORD, 1992, 131 (07) :140-144
[6]   Genetic variation in porcine reproductive and respiratory syndrome virus isolates in the midwestern United States [J].
Kapur, V ;
Elam, MR ;
Pawlovich, TM ;
Murtaugh, MP .
JOURNAL OF GENERAL VIROLOGY, 1996, 77 :1271-1276
[7]   Baculovirus expression and immunological detection of the major structural proteins of porcine reproductive and respiratory syndrome virus [J].
Kreutz, LC ;
Mengeling, WL .
VETERINARY MICROBIOLOGY, 1997, 59 (01) :1-13
[8]   Antibody and cellular immune responses of swine following immunisation with plasmid DNA encoding the PRRS virus ORF's 4, 5, 6 and 7 [J].
Kwang, J ;
Zuckermann, F ;
Ross, G ;
Yang, S ;
Osorio, F ;
Liu, W ;
Low, S .
RESEARCH IN VETERINARY SCIENCE, 1999, 67 (02) :199-201
[9]   Kinetics of humoral immune response to the major structural proteins of the porcine reproductive and respiratory syndrome virus [J].
Loemba, HD ;
Mounir, S ;
Mardassi, H ;
Archambault, D ;
Dea, S .
ARCHIVES OF VIROLOGY, 1996, 141 (3-4) :751-761
[10]   A nested set of six or seven subgenomic mRNAs is formed in cells infected with different isolates of porcine reproductive and respiratory syndrome virus [J].
Meng, XJ ;
Paul, PS ;
Morozov, I ;
Halbur, PG .
JOURNAL OF GENERAL VIROLOGY, 1996, 77 :1265-1270