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Immunological detection of pyrazine-2- carboxylic acid for the detection of pyrazinamide resistance in Mycobacterium tuberculosis

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dc.contributor.author Florentini, E.A.
dc.contributor.author Angulo, N.
dc.contributor.author Gilman, Robert Hugh
dc.contributor.author Alcántara, R.
dc.contributor.author Roncal, E.
dc.contributor.author Antiparra, R.
dc.contributor.author Toscano, E.
dc.contributor.author Vallejos, K.
dc.contributor.author Kirwan, D.
dc.contributor.author Zimic-Peralta, Mirko Juan
dc.contributor.author Sheen Cortavarria, Patricia
dc.date.accessioned 2020-12-14T16:06:23Z
dc.date.available 2020-12-14T16:06:23Z
dc.date.issued 2020
dc.identifier.uri https://hdl.handle.net/20.500.12866/8709
dc.description.abstract Pyrazinamide (PZA) susceptibility testing in Mycobacterium tuberculosis (Mtb) is a current area of development and PZA-resistant strains are increasingly prevalent. Previous studies have demonstrated that the detection of pyrazinoic acid (POA), the metabolite produced by the deamidation of PZA, is a good predictor for PZA resistance since a resistant strain would not convert PZA into POA at a critical required rate, whereas a susceptible strain will do, expelling POA to the extracellular environment at a certain rate, and allowing for quantification of this accumulated analyte. In order to quantify POA, an indirect competitive ELISA (icELISA) test using hyperimmune polyclonal rabbit serum against POA was developed: For this purpose, pure POA was first covalently linked to the highly immunogenic Keyhole Limpet Hemocyanine, and inoculated in rabbits. A construct made of bovine serum albumin (BSA) linked to pure POA and fixed at the bottom of wells was used as a competitor against spiked samples and liquid Mtb culture supernatants. When spiked samples (commercial POA alone) were analyzed, the half maximal inhibitory concentration (IC50) was 1.16 mg/ mL, the limit of detection 200 μg/mL and the assay was specific (it did not detect PZA, IC50 > 20 mg/mL). However, culture supernatants (7H9-OADC-PANTA medium) disrupted the competition and a proper icELISA curve was not obtainable. We consider that, although we have shown that it is feasible to induce antibodies against POA, matrix effects could damage its analytical usefulness; multiple, upcoming ways to solve this obstacle are suggested. en_US
dc.language.iso eng
dc.publisher Public Library of Science
dc.relation.ispartofseries PLoS ONE
dc.rights info:eu-repo/semantics/restrictedAccess
dc.rights.uri https://creativecommons.org/licenses/by-nc-nd/4.0/deed.es
dc.subject Enzyme-linked immunoassays en_US
dc.subject Mycobacterium tuberculosis en_US
dc.subject Antigens en_US
dc.subject Antibodies en_US
dc.subject Immunologic adjuvants en_US
dc.subject Chemical synthesis en_US
dc.subject Tuberculosis en_US
dc.subject Mutation en_US
dc.title Immunological detection of pyrazine-2- carboxylic acid for the detection of pyrazinamide resistance in Mycobacterium tuberculosis en_US
dc.type info:eu-repo/semantics/article
dc.identifier.doi https://doi.org/10.1371/journal.pone.0241600
dc.subject.ocde https://purl.org/pe-repo/ocde/ford#3.02.07
dc.relation.issn 1932-6203


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