TY - JOUR
T1 - Cloning, expression and characterization of immunogenic aminopeptidase N from Brucella melitensis
AU - Contreras-Rodriguez, Araceli
AU - Seleem, Mohamed Naguieb
AU - Schurig, Gerhardt G.
AU - Sriranganathan, Nammalwar
AU - Boyle, Stephen M.
AU - Lopez-Merino, Ahide
PY - 2006/11
Y1 - 2006/11
N2 - A 97-kDa purified aminopeptidase N (PepN) of Brucella melitensis was previously identified to be immunogenic in humans. The B. melitensis pepN gene was cloned, expressed in Escherichia coli and purified by affinity chromatography. The recombinant PepN (rPepN) exhibited the same biochemical properties, specificity and susceptibility to inhibitors as the native PepN. rPepN was evaluated as a diagnostic antigen in an indirect enzyme-linked immunosorbent assay (ELISA) using sera from patients with acute and chronic brucellosis. The specificity of the ELISA was determined with sera from healthy donors. The ELISA had a cutoff value of 0.156 with 100% specificity and 100% sensitivity. Higher sensitivity was obtained using rPepN compared with crude extract from B. melitensis. Anti-PepN sera did not exhibit serological cross-reaction to crude extracts from Rhizobium tropici, Ochrobactrum anthropi, Yersinia enterocolitica 09 or E. coli O157H7.
AB - A 97-kDa purified aminopeptidase N (PepN) of Brucella melitensis was previously identified to be immunogenic in humans. The B. melitensis pepN gene was cloned, expressed in Escherichia coli and purified by affinity chromatography. The recombinant PepN (rPepN) exhibited the same biochemical properties, specificity and susceptibility to inhibitors as the native PepN. rPepN was evaluated as a diagnostic antigen in an indirect enzyme-linked immunosorbent assay (ELISA) using sera from patients with acute and chronic brucellosis. The specificity of the ELISA was determined with sera from healthy donors. The ELISA had a cutoff value of 0.156 with 100% specificity and 100% sensitivity. Higher sensitivity was obtained using rPepN compared with crude extract from B. melitensis. Anti-PepN sera did not exhibit serological cross-reaction to crude extracts from Rhizobium tropici, Ochrobactrum anthropi, Yersinia enterocolitica 09 or E. coli O157H7.
KW - Aminopeptidase N
KW - Brucella
KW - ELISA
KW - PepN
KW - Recombinant
UR - http://www.scopus.com/inward/record.url?scp=33750072588&partnerID=8YFLogxK
U2 - 10.1111/j.1574-695X.2006.00145.x
DO - 10.1111/j.1574-695X.2006.00145.x
M3 - Artículo
C2 - 17064279
SN - 0928-8244
VL - 48
SP - 252
EP - 256
JO - FEMS Immunology and Medical Microbiology
JF - FEMS Immunology and Medical Microbiology
IS - 2
ER -