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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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