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A protein-conjugate approach to develop a monoclonal antibody-based antigen detection test for the diagnosis of human brucellosis

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dc.contributor.author Patra, Kailash P.
dc.contributor.author Saito, Mayuko
dc.contributor.author Atluri, Vidya L.
dc.contributor.author Rolan, Hortensia G.
dc.contributor.author Young, Briana
dc.contributor.author Kerrinnes, Tobias
dc.contributor.author Smits, Henk
dc.contributor.author Ricaldi, Jessica N.
dc.contributor.author Gotuzzo, Eduardo
dc.contributor.author Gilman, Robert H.
dc.contributor.author Tsolis, Renee M.
dc.contributor.author Vinetz, Joseph M.
dc.date.accessioned 2020-06-10T18:12:16Z
dc.date.available 2020-06-10T18:12:16Z
dc.date.issued 2014
dc.identifier.uri https://hdl.handle.net/20.500.12866/8090
dc.description.abstract Human brucellosis is most commonly diagnosed by serology based on agglutination of fixed Brucella abortus as antigen. Nucleic acid amplification techniques have not proven capable of reproducibly and sensitively demonstrating the presence of Brucella DNA in clinical specimens. We sought to optimize a monoclonal antibody-based assay to detect Brucella melitensis lipopolysaccharide in blood by conjugating B. melitensis LPS to keyhole limpet hemocyanin, an immunogenic protein carrier to maximize IgG affinity of monoclonal antibodies. A panel of specific of monoclonal antibodies was obtained that recognized both B. melitensis and B. abortus lipopolysaccharide epitopes. An antigen capture assay was developed that detected B. melitensis in the blood of experimentally infected mice and, in a pilot study, in naturally infected Peruvian subjects. As a proof of principle, a majority (7/10) of the patients with positive blood cultures had B. melitensis lipopolysaccharide detected in the initial blood specimen obtained. One of 10 patients with relapsed brucellosis and negative blood culture had a positive serum antigen test. No seronegative/blood culture negative patients had a positive serum antigen test. Analysis of the pair of monoclonal antibodies (2D1, 2E8) used in the capture ELISA for potential cross-reactivity in the detection of lipopolysaccharides of E. coli O157:H7 and Yersinia enterocolitica O9 showed specificity for Brucella lipopolysaccharide. This new approach to develop antigen-detection monoclonal antibodies against a T cell-independent polysaccharide antigen based on immunogenic protein conjugation may lead to the production of improved rapid point-of-care-deployable assays for the diagnosis of brucellosis and other infectious diseases. en_US
dc.language.iso eng
dc.publisher Public Library of Science
dc.relation.ispartof urn:issn:1935-2735
dc.rights info:eu-repo/semantics/restrictedAccess
dc.rights.uri https://creativecommons.org/licenses/by-nc-nd/4.0/deed.es
dc.subject Antibodies, Bacterial en_US
dc.subject Antibodies, Monoclonal en_US
dc.subject Adult en_US
dc.subject Animals en_US
dc.subject Antigens, Bacterial/blood en_US
dc.subject Brucella abortus/chemistry/isolation & purification en_US
dc.subject Brucella melitensis/chemistry/isolation & purification en_US
dc.subject Brucellosis/diagnosis en_US
dc.subject Enzyme-Linked Immunosorbent Assay/methods en_US
dc.subject Humans en_US
dc.subject Lipopolysaccharides/blood en_US
dc.subject Mice, Inbred BALB C en_US
dc.subject Mice, Inbred C57BL en_US
dc.subject Sensitivity and Specificity en_US
dc.title A protein-conjugate approach to develop a monoclonal antibody-based antigen detection test for the diagnosis of human brucellosis en_US
dc.type info:eu-repo/semantics/article
dc.identifier.doi https://doi.org/10.1371/journal.pntd.0002926
dc.subject.ocde https://purl.org/pe-repo/ocde/ford#3.02.00 es_PE


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