A Multiplex Immunosensor for Detecting Perchlorate-Reducing Bacteria for Environmental Monitoring and Planetary Exploration
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 sur...
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Frontiers Media S.A.
2020-12-01
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Series: | Frontiers in Microbiology |
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Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2020.590736/full |
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author | Ignacio Gallardo-Carreño Mercedes Moreno-Paz Jacobo Aguirre Jacobo Aguirre Jacobo Aguirre Yolanda Blanco Eduardo Alonso-Pintado Isabelle Raymond-Bouchard Catherine Maggiori Luis A. Rivas Luis A. Rivas Anna Engelbrektson Lyle Whyte Víctor Parro |
author_facet | Ignacio Gallardo-Carreño Mercedes Moreno-Paz Jacobo Aguirre Jacobo Aguirre Jacobo Aguirre Yolanda Blanco Eduardo Alonso-Pintado Isabelle Raymond-Bouchard Catherine Maggiori Luis A. Rivas Luis A. Rivas Anna Engelbrektson Lyle Whyte Víctor Parro |
author_sort | Ignacio Gallardo-Carreño |
collection | DOAJ |
description | 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. |
first_indexed | 2024-12-17T04:49:35Z |
format | Article |
id | doaj.art-deb77e5b5255499c8c589168fc1f93e7 |
institution | Directory Open Access Journal |
issn | 1664-302X |
language | English |
last_indexed | 2024-12-17T04:49:35Z |
publishDate | 2020-12-01 |
publisher | Frontiers Media S.A. |
record_format | Article |
series | Frontiers in Microbiology |
spelling | doaj.art-deb77e5b5255499c8c589168fc1f93e72022-12-21T22:02:57ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2020-12-011110.3389/fmicb.2020.590736590736A Multiplex Immunosensor for Detecting Perchlorate-Reducing Bacteria for Environmental Monitoring and Planetary ExplorationIgnacio Gallardo-Carreño0Mercedes Moreno-Paz1Jacobo Aguirre2Jacobo Aguirre3Jacobo Aguirre4Yolanda Blanco5Eduardo Alonso-Pintado6Isabelle Raymond-Bouchard7Catherine Maggiori8Luis A. Rivas9Luis A. Rivas10Anna Engelbrektson11Lyle Whyte12Víctor Parro13Department of Molecular Evolution, Centro de Astrobiología (INTA-CSIC), Madrid, SpainDepartment of Molecular Evolution, Centro de Astrobiología (INTA-CSIC), Madrid, SpainDepartment of Molecular Evolution, Centro de Astrobiología (INTA-CSIC), Madrid, SpainCentro Nacional de Biotecnología, CSIC, Madrid, SpainGrupo Interdisciplinar de Sistemas Complejos (GISC), Madrid, SpainDepartment of Molecular Evolution, Centro de Astrobiología (INTA-CSIC), Madrid, SpainDepartment of Molecular Evolution, Centro de Astrobiología (INTA-CSIC), Madrid, SpainDepartment of Natural Resource Sciences, McGill University, Sainte-Anne-de-Bellevue, QC, CanadaDepartment of Natural Resource Sciences, McGill University, Sainte-Anne-de-Bellevue, QC, CanadaDepartment of Molecular Evolution, Centro de Astrobiología (INTA-CSIC), Madrid, SpainInmunología y Genética Aplicada, S.A. (INGENASA), Madrid, SpainDepartment of Plant & Microbial Biology, University of California, Berkeley, Berkeley, CA, United StatesDepartment of Natural Resource Sciences, McGill University, Sainte-Anne-de-Bellevue, QC, CanadaDepartment of Molecular Evolution, Centro de Astrobiología (INTA-CSIC), Madrid, SpainPerchlorate 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.https://www.frontiersin.org/articles/10.3389/fmicb.2020.590736/fullperchlorateperchlorate-reducing bacteriaantibody microarraysbiochiplife detectionplanetary exploration |
spellingShingle | Ignacio Gallardo-Carreño Mercedes Moreno-Paz Jacobo Aguirre Jacobo Aguirre Jacobo Aguirre Yolanda Blanco Eduardo Alonso-Pintado Isabelle Raymond-Bouchard Catherine Maggiori Luis A. Rivas Luis A. Rivas Anna Engelbrektson Lyle Whyte Víctor Parro A Multiplex Immunosensor for Detecting Perchlorate-Reducing Bacteria for Environmental Monitoring and Planetary Exploration Frontiers in Microbiology perchlorate perchlorate-reducing bacteria antibody microarrays biochip life detection planetary exploration |
title | 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_short | 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 |
topic | perchlorate perchlorate-reducing bacteria antibody microarrays biochip life detection planetary exploration |
url | https://www.frontiersin.org/articles/10.3389/fmicb.2020.590736/full |
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