A Multiplex Immunosensor for Detecting Perchlorate-Reducing Bacteria for Environmental Monitoring and Planetary Exploration
© 2020 Gallardo-Carreño, Moreno-Paz, Aguirre, Blanco, Alonso-Pintado, Raymond-Bouchard, Maggiori, Rivas, Engelbrektson, Whyte and Parro. Perchlorate anions are produced by chemical industries and are important contaminants in certain natural ecosystems. Perchlorate also occurs in some natural and un...
Published in: | Frontiers in Microbiology |
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Main Authors: | , , , , , , , , , , |
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Frontiers Media
2020
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Online Access: | http://hdl.handle.net/10261/230083 https://doi.org/10.3389/fmicb.2020.590736 https://doi.org/10.13039/501100011033 https://doi.org/10.13039/501100003329 |
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Digital.CSIC (Spanish National Research Council) |
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topic |
Perchlorate Perchlorate-reducing bacteria Antibody microarrays Biochip Life detection Planetary exploration Mars Graph theory |
spellingShingle |
Perchlorate Perchlorate-reducing bacteria Antibody microarrays Biochip Life detection Planetary exploration Mars Graph theory Gallardo-Carreño, Ignacio Moreno-Paz, Mercedes Aguirre, Jacobo Blanco, Yolanda Alonso-Pintado, Eduardo Raymond-Bouchard, Isabelle Maggiori, Catherine Rivas, Luis A. Engelbrektson, Anna Whyte, Lyle Parro-García, Víctor A Multiplex Immunosensor for Detecting Perchlorate-Reducing Bacteria for Environmental Monitoring and Planetary Exploration |
topic_facet |
Perchlorate Perchlorate-reducing bacteria Antibody microarrays Biochip Life detection Planetary exploration Mars Graph theory |
description |
© 2020 Gallardo-Carreño, Moreno-Paz, Aguirre, Blanco, Alonso-Pintado, Raymond-Bouchard, Maggiori, Rivas, Engelbrektson, Whyte and Parro. Perchlorate anions are produced by chemical industries and are important contaminants in certain natural ecosystems. Perchlorate also occurs in some natural and uncontaminated environments such as the Atacama Desert, the high Arctic or the Antarctic Dry Valleys, and is especially abundant on the surface of Mars. As some bacterial strains are capable of using perchlorate as an electron acceptor under anaerobic conditions, their detection is relevant for environmental monitoring on Earth as well as for the search for life on Mars. We have developed an antibody microarray with 20 polyclonal antibodies to detect perchlorate-reducing bacteria (PRB) strains and two crucial and highly conserved enzymes involved in perchlorate respiration: perchlorate reductase and chlorite dismutase. We determined the cross-reactivity, the working concentration, and the limit of detection of each antibody individually and in a multiplex format by Fluorescent Sandwich Microarray Immunoassay. Although most of them exhibited relatively high sensitivity and specificity, we applied a deconvolution method based on graph theory to discriminate between specific signals and cross-reactions from related microorganisms. We validated the system by analyzing multiple bacterial isolates, crude extracts from contaminated reactors and salt-rich natural samples from the high Arctic. The PRB detecting chip (PRBCHIP) allowed us to detect and classify environmental isolates as well as to detect similar strains by using crude extracts obtained from 0.5 g even from soils with low organic-matter levels (<103 cells/g of soil). Our results demonstrated that PRBCHIP is a valuable tool for sensitive and reliable detection of perchlorate-reducing bacteria for research purposes, environmental monitoring and planetary exploration. This work was funded by the Ministry of Economy and Competitiveness, (MINECO)/Fondo Europeo de ... |
author2 |
Ministerio de Economía y Competitividad (España) Ministerio de Ciencia, Innovación y Universidades (España) Agencia Estatal de Investigación (España) |
format |
Article in Journal/Newspaper |
author |
Gallardo-Carreño, Ignacio Moreno-Paz, Mercedes Aguirre, Jacobo Blanco, Yolanda Alonso-Pintado, Eduardo Raymond-Bouchard, Isabelle Maggiori, Catherine Rivas, Luis A. Engelbrektson, Anna Whyte, Lyle Parro-García, Víctor |
author_facet |
Gallardo-Carreño, Ignacio Moreno-Paz, Mercedes Aguirre, Jacobo Blanco, Yolanda Alonso-Pintado, Eduardo Raymond-Bouchard, Isabelle Maggiori, Catherine Rivas, Luis A. Engelbrektson, Anna Whyte, Lyle Parro-García, Víctor |
author_sort |
Gallardo-Carreño, Ignacio |
title |
A Multiplex Immunosensor for Detecting Perchlorate-Reducing Bacteria for Environmental Monitoring and Planetary Exploration |
title_short |
A Multiplex Immunosensor for Detecting Perchlorate-Reducing Bacteria for Environmental Monitoring and Planetary Exploration |
title_full |
A Multiplex Immunosensor for Detecting Perchlorate-Reducing Bacteria for Environmental Monitoring and Planetary Exploration |
title_fullStr |
A Multiplex Immunosensor for Detecting Perchlorate-Reducing Bacteria for Environmental Monitoring and Planetary Exploration |
title_full_unstemmed |
A Multiplex Immunosensor for Detecting Perchlorate-Reducing Bacteria for Environmental Monitoring and Planetary Exploration |
title_sort |
multiplex immunosensor for detecting perchlorate-reducing bacteria for environmental monitoring and planetary exploration |
publisher |
Frontiers Media |
publishDate |
2020 |
url |
http://hdl.handle.net/10261/230083 https://doi.org/10.3389/fmicb.2020.590736 https://doi.org/10.13039/501100011033 https://doi.org/10.13039/501100003329 |
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ENVELOPE(-57.933,-57.933,-63.317,-63.317) ENVELOPE(-55.233,-55.233,-61.250,-61.250) ENVELOPE(-57.333,-57.333,-64.200,-64.200) ENVELOPE(-62.300,-62.300,-64.083,-64.083) |
geographic |
Aguirre Antarctic Arctic Blanco Bouchard Moreno The Antarctic |
geographic_facet |
Aguirre Antarctic Arctic Blanco Bouchard Moreno The Antarctic |
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Antarc* Antarctic Arctic Pintado |
genre_facet |
Antarc* Antarctic Arctic Pintado |
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https://doi.org/10.3389/fmicb.2020.59073610.13039/50110001103310.13039/501100003329 |
container_title |
Frontiers in Microbiology |
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11 |
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ftcsic:oai:digital.csic.es:10261/230083 2024-02-11T09:58:45+01:00 A Multiplex Immunosensor for Detecting Perchlorate-Reducing Bacteria for Environmental Monitoring and Planetary Exploration Gallardo-Carreño, Ignacio Moreno-Paz, Mercedes Aguirre, Jacobo Blanco, Yolanda Alonso-Pintado, Eduardo Raymond-Bouchard, Isabelle Maggiori, Catherine Rivas, Luis A. Engelbrektson, Anna Whyte, Lyle Parro-García, Víctor Ministerio de Economía y Competitividad (España) Ministerio de Ciencia, Innovación y Universidades (España) Agencia Estatal de Investigación (España) 2020-12 http://hdl.handle.net/10261/230083 https://doi.org/10.3389/fmicb.2020.590736 https://doi.org/10.13039/501100011033 https://doi.org/10.13039/501100003329 unknown Frontiers Media #PLACEHOLDER_PARENT_METADATA_VALUE# info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/ESP2015-69540-R info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/RTI2018-094368-B-I00 RTI2018-094368-B-I00/AEI/10.13039/501100011033 info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/MDM-2017-0737 info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/FIS2017-89773-P info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/SEV2017-0712-IG-C Publisher's version http://dx.doi.org/10.3389/fmicb.2020.590736 Sí Frontiers in Microbiology 11: 590736 (2020) http://hdl.handle.net/10261/230083 doi:10.3389/fmicb.2020.590736 1664-302X http://dx.doi.org/10.13039/501100011033 http://dx.doi.org/10.13039/501100003329 33391207 open Perchlorate Perchlorate-reducing bacteria Antibody microarrays Biochip Life detection Planetary exploration Mars Graph theory artículo http://purl.org/coar/resource_type/c_6501 2020 ftcsic https://doi.org/10.3389/fmicb.2020.59073610.13039/50110001103310.13039/501100003329 2024-01-16T11:03:02Z © 2020 Gallardo-Carreño, Moreno-Paz, Aguirre, Blanco, Alonso-Pintado, Raymond-Bouchard, Maggiori, Rivas, Engelbrektson, Whyte and Parro. Perchlorate anions are produced by chemical industries and are important contaminants in certain natural ecosystems. Perchlorate also occurs in some natural and uncontaminated environments such as the Atacama Desert, the high Arctic or the Antarctic Dry Valleys, and is especially abundant on the surface of Mars. As some bacterial strains are capable of using perchlorate as an electron acceptor under anaerobic conditions, their detection is relevant for environmental monitoring on Earth as well as for the search for life on Mars. We have developed an antibody microarray with 20 polyclonal antibodies to detect perchlorate-reducing bacteria (PRB) strains and two crucial and highly conserved enzymes involved in perchlorate respiration: perchlorate reductase and chlorite dismutase. We determined the cross-reactivity, the working concentration, and the limit of detection of each antibody individually and in a multiplex format by Fluorescent Sandwich Microarray Immunoassay. Although most of them exhibited relatively high sensitivity and specificity, we applied a deconvolution method based on graph theory to discriminate between specific signals and cross-reactions from related microorganisms. We validated the system by analyzing multiple bacterial isolates, crude extracts from contaminated reactors and salt-rich natural samples from the high Arctic. The PRB detecting chip (PRBCHIP) allowed us to detect and classify environmental isolates as well as to detect similar strains by using crude extracts obtained from 0.5 g even from soils with low organic-matter levels (<103 cells/g of soil). Our results demonstrated that PRBCHIP is a valuable tool for sensitive and reliable detection of perchlorate-reducing bacteria for research purposes, environmental monitoring and planetary exploration. This work was funded by the Ministry of Economy and Competitiveness, (MINECO)/Fondo Europeo de ... Article in Journal/Newspaper Antarc* Antarctic Arctic Pintado Digital.CSIC (Spanish National Research Council) Aguirre ENVELOPE(-57.933,-57.933,-63.317,-63.317) Antarctic Arctic Blanco ENVELOPE(-55.233,-55.233,-61.250,-61.250) Bouchard ENVELOPE(-57.333,-57.333,-64.200,-64.200) Moreno ENVELOPE(-62.300,-62.300,-64.083,-64.083) The Antarctic Frontiers in Microbiology 11 |