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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Main Authors: Ignacio Gallardo-Carreño, Mercedes Moreno-Paz, Jacobo Aguirre, Yolanda Blanco, Eduardo Alonso-Pintado, Isabelle Raymond-Bouchard, Catherine Maggiori, Luis A. Rivas, Anna Engelbrektson, Lyle Whyte, Víctor Parro
Format: Article
Language:English
Published: Frontiers Media S.A. 2020-12-01
Series:Frontiers in Microbiology
Subjects:
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.
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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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