Immunological detection of pyrazine-2-carboxylic acid for the detection of pyrazinamide resistance in Mycobacterium tuberculosis
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...
Gespeichert in:
| Veröffentlicht in: | PloS one 2020-11, Vol.15 (11), p.e0241600 |
|---|---|
| Hauptverfasser: | , , , , , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | |
|---|---|
| container_issue | 11 |
| container_start_page | e0241600 |
| container_title | PloS one |
| container_volume | 15 |
| creator | Florentini, Edgar A Angulo, Noelia Gilman, Robert H Alcántara, Roberto Roncal, Elisa Antiparra, Ricardo Toscano, Emily Vallejos, Katherine Kirwan, Danni Zimic, Mirko Sheen, Patricia |
| description | 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. |
| doi_str_mv | 10.1371/journal.pone.0241600 |
| format | Article |
| fullrecord | <record><control><sourceid>gale_plos_</sourceid><recordid>TN_cdi_plos_journals_2457961835</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><galeid>A640707122</galeid><doaj_id>oai_doaj_org_article_323a61b43f9241a1b4b39d221ba6862c</doaj_id><sourcerecordid>A640707122</sourcerecordid><originalsourceid>FETCH-LOGICAL-c692t-a7bccc5408dc0d7a884e08fdd11a5b1aa34503f9735f817ca1c38630fa0a3b423</originalsourceid><addsrcrecordid>eNqNkl2L1DAUhoso7rr6D0QLguBFx3y0aXsjLIsfAysLft2G05N0JkPbzCap7HjlTzfjdJcprCC5SDh5znuSlzdJnlOyoLykbzd2dAN0i60d9IKwnApCHiSntOYsE4zwh0fnk-SJ9xtCCl4J8Tg54ZwWtK6K0-T3su_HwXZ2ZRC6VOmgMRg7pLZNtzsHv8ygM5YhuMbe7DqDKaBRaWtdGtb6Xh56o3TqtDc-wIA6NUP6eYe2AQzambFPw9hoh2NnI_I0edRC5_WzaT9Lvn94_-3iU3Z59XF5cX6ZoahZyKBsELHISaWQqBKqKtekapWiFIqGAvC8ILytS160FS0RKMa_ctICAd7kjJ8lLw-62zhWTuZ5yfKirAWteBGJ5YFQFjZy60wPbictGPm3YN1KggsGOy054yBok8eB0XiIp4bXijHagKgEw6j1bpo2Nr1WqIfgoJuJzm8Gs5Yr-1OWIud1nUeBV5OAs9ej9uEfT56oFcRXmaG1UQx741Gei5yUpKRs__XFPVRcSvcGY3xaE-uzhjezhsgEfRNWMHovl1-__D979WPOvj5i1xq6sPa2G_cB8nMwP4DorPdOt3fOUSL36b91Q-7TL6f0x7YXx67fNd3Gnf8BZkEB-w</addsrcrecordid><sourcetype>Open Website</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2457961835</pqid></control><display><type>article</type><title>Immunological detection of pyrazine-2-carboxylic acid for the detection of pyrazinamide resistance in Mycobacterium tuberculosis</title><source>MEDLINE</source><source>DOAJ Directory of Open Access Journals</source><source>Public Library of Science (PLoS) Journals Open Access</source><source>EZB-FREE-00999 freely available EZB journals</source><source>PubMed Central</source><source>Free Full-Text Journals in Chemistry</source><creator>Florentini, Edgar A ; Angulo, Noelia ; Gilman, Robert H ; Alcántara, Roberto ; Roncal, Elisa ; Antiparra, Ricardo ; Toscano, Emily ; Vallejos, Katherine ; Kirwan, Danni ; Zimic, Mirko ; Sheen, Patricia</creator><contributor>D'Auria, Sabato</contributor><creatorcontrib>Florentini, Edgar A ; Angulo, Noelia ; Gilman, Robert H ; Alcántara, Roberto ; Roncal, Elisa ; Antiparra, Ricardo ; Toscano, Emily ; Vallejos, Katherine ; Kirwan, Danni ; Zimic, Mirko ; Sheen, Patricia ; D'Auria, Sabato</creatorcontrib><description>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.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0241600</identifier><identifier>PMID: 33151985</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Animals ; Antibodies ; Antibodies - chemistry ; Antibodies - immunology ; Antitubercular Agents - toxicity ; Biology and Life Sciences ; Bovine serum albumin ; Carboxylic acids ; Cytoplasm ; Drug resistance ; Drug Resistance, Bacterial ; Enzyme-linked immunosorbent assay ; Enzyme-Linked Immunosorbent Assay - methods ; Health aspects ; Immune response ; Immunoconjugates - chemistry ; Immunoconjugates - immunology ; Immunogenicity ; Immunology ; Inhibitory Concentration 50 ; Medical research ; Medicine and Health Sciences ; Metabolites ; Microbial drug resistance ; Mortality ; Mutation ; Mycobacterium tuberculosis ; Mycobacterium tuberculosis - drug effects ; Observations ; Patient outcomes ; Pyrazinamide ; Pyrazinamide - analogs & derivatives ; Pyrazinamide - chemistry ; Pyrazinamide - immunology ; Pyrazinamide - toxicity ; Pyrazine ; Rabbits ; Research and Analysis Methods ; Serum albumin ; Serum Albumin, Bovine - chemistry ; Testing ; Toxicity Tests - methods ; Tuberculosis</subject><ispartof>PloS one, 2020-11, Vol.15 (11), p.e0241600</ispartof><rights>COPYRIGHT 2020 Public Library of Science</rights><rights>2020 Florentini et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2020 Florentini et al 2020 Florentini et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c692t-a7bccc5408dc0d7a884e08fdd11a5b1aa34503f9735f817ca1c38630fa0a3b423</citedby><cites>FETCH-LOGICAL-c692t-a7bccc5408dc0d7a884e08fdd11a5b1aa34503f9735f817ca1c38630fa0a3b423</cites><orcidid>0000-0002-7118-9301 ; 0000-0002-2434-5244</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC7643994/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC7643994/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,2102,2928,23866,27924,27925,53791,53793,79600,79601</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/33151985$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>D'Auria, Sabato</contributor><creatorcontrib>Florentini, Edgar A</creatorcontrib><creatorcontrib>Angulo, Noelia</creatorcontrib><creatorcontrib>Gilman, Robert H</creatorcontrib><creatorcontrib>Alcántara, Roberto</creatorcontrib><creatorcontrib>Roncal, Elisa</creatorcontrib><creatorcontrib>Antiparra, Ricardo</creatorcontrib><creatorcontrib>Toscano, Emily</creatorcontrib><creatorcontrib>Vallejos, Katherine</creatorcontrib><creatorcontrib>Kirwan, Danni</creatorcontrib><creatorcontrib>Zimic, Mirko</creatorcontrib><creatorcontrib>Sheen, Patricia</creatorcontrib><title>Immunological detection of pyrazine-2-carboxylic acid for the detection of pyrazinamide resistance in Mycobacterium tuberculosis</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>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.</description><subject>Animals</subject><subject>Antibodies</subject><subject>Antibodies - chemistry</subject><subject>Antibodies - immunology</subject><subject>Antitubercular Agents - toxicity</subject><subject>Biology and Life Sciences</subject><subject>Bovine serum albumin</subject><subject>Carboxylic acids</subject><subject>Cytoplasm</subject><subject>Drug resistance</subject><subject>Drug Resistance, Bacterial</subject><subject>Enzyme-linked immunosorbent assay</subject><subject>Enzyme-Linked Immunosorbent Assay - methods</subject><subject>Health aspects</subject><subject>Immune response</subject><subject>Immunoconjugates - chemistry</subject><subject>Immunoconjugates - immunology</subject><subject>Immunogenicity</subject><subject>Immunology</subject><subject>Inhibitory Concentration 50</subject><subject>Medical research</subject><subject>Medicine and Health Sciences</subject><subject>Metabolites</subject><subject>Microbial drug resistance</subject><subject>Mortality</subject><subject>Mutation</subject><subject>Mycobacterium tuberculosis</subject><subject>Mycobacterium tuberculosis - drug effects</subject><subject>Observations</subject><subject>Patient outcomes</subject><subject>Pyrazinamide</subject><subject>Pyrazinamide - analogs & derivatives</subject><subject>Pyrazinamide - chemistry</subject><subject>Pyrazinamide - immunology</subject><subject>Pyrazinamide - toxicity</subject><subject>Pyrazine</subject><subject>Rabbits</subject><subject>Research and Analysis Methods</subject><subject>Serum albumin</subject><subject>Serum Albumin, Bovine - chemistry</subject><subject>Testing</subject><subject>Toxicity Tests - methods</subject><subject>Tuberculosis</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><sourceid>DOA</sourceid><recordid>eNqNkl2L1DAUhoso7rr6D0QLguBFx3y0aXsjLIsfAysLft2G05N0JkPbzCap7HjlTzfjdJcprCC5SDh5znuSlzdJnlOyoLykbzd2dAN0i60d9IKwnApCHiSntOYsE4zwh0fnk-SJ9xtCCl4J8Tg54ZwWtK6K0-T3su_HwXZ2ZRC6VOmgMRg7pLZNtzsHv8ygM5YhuMbe7DqDKaBRaWtdGtb6Xh56o3TqtDc-wIA6NUP6eYe2AQzambFPw9hoh2NnI_I0edRC5_WzaT9Lvn94_-3iU3Z59XF5cX6ZoahZyKBsELHISaWQqBKqKtekapWiFIqGAvC8ILytS160FS0RKMa_ctICAd7kjJ8lLw-62zhWTuZ5yfKirAWteBGJ5YFQFjZy60wPbictGPm3YN1KggsGOy054yBok8eB0XiIp4bXijHagKgEw6j1bpo2Nr1WqIfgoJuJzm8Gs5Yr-1OWIud1nUeBV5OAs9ej9uEfT56oFcRXmaG1UQx741Gei5yUpKRs__XFPVRcSvcGY3xaE-uzhjezhsgEfRNWMHovl1-__D979WPOvj5i1xq6sPa2G_cB8nMwP4DorPdOt3fOUSL36b91Q-7TL6f0x7YXx67fNd3Gnf8BZkEB-w</recordid><startdate>20201105</startdate><enddate>20201105</enddate><creator>Florentini, Edgar A</creator><creator>Angulo, Noelia</creator><creator>Gilman, Robert H</creator><creator>Alcántara, Roberto</creator><creator>Roncal, Elisa</creator><creator>Antiparra, Ricardo</creator><creator>Toscano, Emily</creator><creator>Vallejos, Katherine</creator><creator>Kirwan, Danni</creator><creator>Zimic, Mirko</creator><creator>Sheen, Patricia</creator><general>Public Library of Science</general><general>Public Library of Science (PLoS)</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>IOV</scope><scope>ISR</scope><scope>3V.</scope><scope>7QG</scope><scope>7QL</scope><scope>7QO</scope><scope>7RV</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TG</scope><scope>7TM</scope><scope>7U9</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>KB0</scope><scope>KL.</scope><scope>L6V</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>M7S</scope><scope>NAPCQ</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PATMY</scope><scope>PDBOC</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>PTHSS</scope><scope>PYCSY</scope><scope>RC3</scope><scope>5PM</scope><scope>DOA</scope><orcidid>https://orcid.org/0000-0002-7118-9301</orcidid><orcidid>https://orcid.org/0000-0002-2434-5244</orcidid></search><sort><creationdate>20201105</creationdate><title>Immunological detection of pyrazine-2-carboxylic acid for the detection of pyrazinamide resistance in Mycobacterium tuberculosis</title><author>Florentini, Edgar A ; Angulo, Noelia ; Gilman, Robert H ; Alcántara, Roberto ; Roncal, Elisa ; Antiparra, Ricardo ; Toscano, Emily ; Vallejos, Katherine ; Kirwan, Danni ; Zimic, Mirko ; Sheen, Patricia</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c692t-a7bccc5408dc0d7a884e08fdd11a5b1aa34503f9735f817ca1c38630fa0a3b423</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2020</creationdate><topic>Animals</topic><topic>Antibodies</topic><topic>Antibodies - chemistry</topic><topic>Antibodies - immunology</topic><topic>Antitubercular Agents - toxicity</topic><topic>Biology and Life Sciences</topic><topic>Bovine serum albumin</topic><topic>Carboxylic acids</topic><topic>Cytoplasm</topic><topic>Drug resistance</topic><topic>Drug Resistance, Bacterial</topic><topic>Enzyme-linked immunosorbent assay</topic><topic>Enzyme-Linked Immunosorbent Assay - methods</topic><topic>Health aspects</topic><topic>Immune response</topic><topic>Immunoconjugates - chemistry</topic><topic>Immunoconjugates - immunology</topic><topic>Immunogenicity</topic><topic>Immunology</topic><topic>Inhibitory Concentration 50</topic><topic>Medical research</topic><topic>Medicine and Health Sciences</topic><topic>Metabolites</topic><topic>Microbial drug resistance</topic><topic>Mortality</topic><topic>Mutation</topic><topic>Mycobacterium tuberculosis</topic><topic>Mycobacterium tuberculosis - drug effects</topic><topic>Observations</topic><topic>Patient outcomes</topic><topic>Pyrazinamide</topic><topic>Pyrazinamide - analogs & derivatives</topic><topic>Pyrazinamide - chemistry</topic><topic>Pyrazinamide - immunology</topic><topic>Pyrazinamide - toxicity</topic><topic>Pyrazine</topic><topic>Rabbits</topic><topic>Research and Analysis Methods</topic><topic>Serum albumin</topic><topic>Serum Albumin, Bovine - chemistry</topic><topic>Testing</topic><topic>Toxicity Tests - methods</topic><topic>Tuberculosis</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Florentini, Edgar A</creatorcontrib><creatorcontrib>Angulo, Noelia</creatorcontrib><creatorcontrib>Gilman, Robert H</creatorcontrib><creatorcontrib>Alcántara, Roberto</creatorcontrib><creatorcontrib>Roncal, Elisa</creatorcontrib><creatorcontrib>Antiparra, Ricardo</creatorcontrib><creatorcontrib>Toscano, Emily</creatorcontrib><creatorcontrib>Vallejos, Katherine</creatorcontrib><creatorcontrib>Kirwan, Danni</creatorcontrib><creatorcontrib>Zimic, Mirko</creatorcontrib><creatorcontrib>Sheen, Patricia</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Gale In Context: Opposing Viewpoints</collection><collection>Gale In Context: Science</collection><collection>ProQuest Central (Corporate)</collection><collection>Animal Behavior Abstracts</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Nursing & Allied Health Database</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Immunology Abstracts</collection><collection>Meteorological & Geoastrophysical Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Agricultural Science Collection</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Public Health Database</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>Materials Science & Engineering Collection</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>Advanced Technologies & Aerospace Collection</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Technology Collection</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Materials Science Collection</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Materials Science Database</collection><collection>Nursing & Allied Health Database (Alumni Edition)</collection><collection>Meteorological & Geoastrophysical Abstracts - Academic</collection><collection>ProQuest Engineering Collection</collection><collection>ProQuest Biological Science Collection</collection><collection>Agricultural Science Database</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biological Science Database</collection><collection>Engineering Database</collection><collection>Nursing & Allied Health Premium</collection><collection>Advanced Technologies & Aerospace Database</collection><collection>ProQuest Advanced Technologies & Aerospace Collection</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Environmental Science Database</collection><collection>Materials Science Collection</collection><collection>Publicly Available Content Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>Engineering Collection</collection><collection>Environmental Science Collection</collection><collection>Genetics Abstracts</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Florentini, Edgar A</au><au>Angulo, Noelia</au><au>Gilman, Robert H</au><au>Alcántara, Roberto</au><au>Roncal, Elisa</au><au>Antiparra, Ricardo</au><au>Toscano, Emily</au><au>Vallejos, Katherine</au><au>Kirwan, Danni</au><au>Zimic, Mirko</au><au>Sheen, Patricia</au><au>D'Auria, Sabato</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Immunological detection of pyrazine-2-carboxylic acid for the detection of pyrazinamide resistance in Mycobacterium tuberculosis</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2020-11-05</date><risdate>2020</risdate><volume>15</volume><issue>11</issue><spage>e0241600</spage><pages>e0241600-</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><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.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>33151985</pmid><doi>10.1371/journal.pone.0241600</doi><orcidid>https://orcid.org/0000-0002-7118-9301</orcidid><orcidid>https://orcid.org/0000-0002-2434-5244</orcidid><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1932-6203 |
| ispartof | PloS one, 2020-11, Vol.15 (11), p.e0241600 |
| issn | 1932-6203 1932-6203 |
| language | eng |
| recordid | cdi_plos_journals_2457961835 |
| source | MEDLINE; DOAJ Directory of Open Access Journals; Public Library of Science (PLoS) Journals Open Access; EZB-FREE-00999 freely available EZB journals; PubMed Central; Free Full-Text Journals in Chemistry |
| subjects | Animals Antibodies Antibodies - chemistry Antibodies - immunology Antitubercular Agents - toxicity Biology and Life Sciences Bovine serum albumin Carboxylic acids Cytoplasm Drug resistance Drug Resistance, Bacterial Enzyme-linked immunosorbent assay Enzyme-Linked Immunosorbent Assay - methods Health aspects Immune response Immunoconjugates - chemistry Immunoconjugates - immunology Immunogenicity Immunology Inhibitory Concentration 50 Medical research Medicine and Health Sciences Metabolites Microbial drug resistance Mortality Mutation Mycobacterium tuberculosis Mycobacterium tuberculosis - drug effects Observations Patient outcomes Pyrazinamide Pyrazinamide - analogs & derivatives Pyrazinamide - chemistry Pyrazinamide - immunology Pyrazinamide - toxicity Pyrazine Rabbits Research and Analysis Methods Serum albumin Serum Albumin, Bovine - chemistry Testing Toxicity Tests - methods Tuberculosis |
| title | Immunological detection of pyrazine-2-carboxylic acid for the detection of pyrazinamide resistance in Mycobacterium tuberculosis |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-29T04%3A49%3A02IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Immunological%20detection%20of%20pyrazine-2-carboxylic%20acid%20for%20the%20detection%20of%20pyrazinamide%20resistance%20in%20Mycobacterium%20tuberculosis&rft.jtitle=PloS%20one&rft.au=Florentini,%20Edgar%20A&rft.date=2020-11-05&rft.volume=15&rft.issue=11&rft.spage=e0241600&rft.pages=e0241600-&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0241600&rft_dat=%3Cgale_plos_%3EA640707122%3C/gale_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=2457961835&rft_id=info:pmid/33151985&rft_galeid=A640707122&rft_doaj_id=oai_doaj_org_article_323a61b43f9241a1b4b39d221ba6862c&rfr_iscdi=true |