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Detection and differentiation of Burkholderia species with pathogenic potential in environmental soil samples

The Burkholderia pseudomallei phylogenetic cluster includes B. pseudomallei, B. mallei, B. thailandensis, B. oklahomensis, B. humptydooensis and B. singularis. Regarded as the only pathogenic members of this group, B. pseudomallei and B. mallei cause the diseases melioidosis and glanders, respective...

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Autores principales: Janesomboon, Sujintana, Muangsombut, Veerachat, Srinon, Varintip, Meethai, Chatruthai, Tharinjaroen, Chayada S., Amornchai, Premjit, Withatanung, Patoo, Chantratita, Narisara, Mayo, Mark, Wuthiekanun, Vanaporn, Currie, Bart J., Stevens, Joanne M., Korbsrisate, Sunee
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7790303/
https://www.ncbi.nlm.nih.gov/pubmed/33411797
http://dx.doi.org/10.1371/journal.pone.0245175
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author Janesomboon, Sujintana
Muangsombut, Veerachat
Srinon, Varintip
Meethai, Chatruthai
Tharinjaroen, Chayada S.
Amornchai, Premjit
Withatanung, Patoo
Chantratita, Narisara
Mayo, Mark
Wuthiekanun, Vanaporn
Currie, Bart J.
Stevens, Joanne M.
Korbsrisate, Sunee
author_facet Janesomboon, Sujintana
Muangsombut, Veerachat
Srinon, Varintip
Meethai, Chatruthai
Tharinjaroen, Chayada S.
Amornchai, Premjit
Withatanung, Patoo
Chantratita, Narisara
Mayo, Mark
Wuthiekanun, Vanaporn
Currie, Bart J.
Stevens, Joanne M.
Korbsrisate, Sunee
author_sort Janesomboon, Sujintana
collection PubMed
description The Burkholderia pseudomallei phylogenetic cluster includes B. pseudomallei, B. mallei, B. thailandensis, B. oklahomensis, B. humptydooensis and B. singularis. Regarded as the only pathogenic members of this group, B. pseudomallei and B. mallei cause the diseases melioidosis and glanders, respectively. Additionally, variant strains of B. pseudomallei and B. thailandensis exist that include the geographically restricted B. pseudomallei that express a B. mallei-like BimA protein (BPBM), and B. thailandensis that express a B. pseudomallei-like capsular polysaccharide (BTCV). To establish a PCR-based assay for the detection of pathogenic Burkholderia species or their variants, five PCR primers were designed to amplify species-specific sequences within the bimA (Burkholderia intracellular motility A) gene. Our multiplex PCR assay could distinguish pathogenic B. pseudomallei and BPBM from the non-pathogenic B. thailandensis and the BTCV strains. A second singleplex PCR successfully discriminated the BTCV from B. thailandensis. Apart from B. humptydooensis, specificity testing against other Burkholderia spp., as well as other Gram-negative and Gram-positive bacteria produced a negative result. The detection limit of the multiplex PCR in soil samples artificially spiked with known quantities of B. pseudomallei and B. thailandensis were 5 and 6 CFU/g soil, respectively. Furthermore, comparison between standard bacterial culture and the multiplex PCR to detect B. pseudomallei from 34 soil samples, collected from an endemic area of melioidosis, showed high sensitivity and specificity. This robust, sensitive, and specific PCR assay will be a useful tool for epidemiological study of B. pseudomallei and closely related members with pathogenic potential in soil.
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spelling pubmed-77903032021-01-27 Detection and differentiation of Burkholderia species with pathogenic potential in environmental soil samples Janesomboon, Sujintana Muangsombut, Veerachat Srinon, Varintip Meethai, Chatruthai Tharinjaroen, Chayada S. Amornchai, Premjit Withatanung, Patoo Chantratita, Narisara Mayo, Mark Wuthiekanun, Vanaporn Currie, Bart J. Stevens, Joanne M. Korbsrisate, Sunee PLoS One Research Article The Burkholderia pseudomallei phylogenetic cluster includes B. pseudomallei, B. mallei, B. thailandensis, B. oklahomensis, B. humptydooensis and B. singularis. Regarded as the only pathogenic members of this group, B. pseudomallei and B. mallei cause the diseases melioidosis and glanders, respectively. Additionally, variant strains of B. pseudomallei and B. thailandensis exist that include the geographically restricted B. pseudomallei that express a B. mallei-like BimA protein (BPBM), and B. thailandensis that express a B. pseudomallei-like capsular polysaccharide (BTCV). To establish a PCR-based assay for the detection of pathogenic Burkholderia species or their variants, five PCR primers were designed to amplify species-specific sequences within the bimA (Burkholderia intracellular motility A) gene. Our multiplex PCR assay could distinguish pathogenic B. pseudomallei and BPBM from the non-pathogenic B. thailandensis and the BTCV strains. A second singleplex PCR successfully discriminated the BTCV from B. thailandensis. Apart from B. humptydooensis, specificity testing against other Burkholderia spp., as well as other Gram-negative and Gram-positive bacteria produced a negative result. The detection limit of the multiplex PCR in soil samples artificially spiked with known quantities of B. pseudomallei and B. thailandensis were 5 and 6 CFU/g soil, respectively. Furthermore, comparison between standard bacterial culture and the multiplex PCR to detect B. pseudomallei from 34 soil samples, collected from an endemic area of melioidosis, showed high sensitivity and specificity. This robust, sensitive, and specific PCR assay will be a useful tool for epidemiological study of B. pseudomallei and closely related members with pathogenic potential in soil. Public Library of Science 2021-01-07 /pmc/articles/PMC7790303/ /pubmed/33411797 http://dx.doi.org/10.1371/journal.pone.0245175 Text en © 2021 Janesomboon et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Janesomboon, Sujintana
Muangsombut, Veerachat
Srinon, Varintip
Meethai, Chatruthai
Tharinjaroen, Chayada S.
Amornchai, Premjit
Withatanung, Patoo
Chantratita, Narisara
Mayo, Mark
Wuthiekanun, Vanaporn
Currie, Bart J.
Stevens, Joanne M.
Korbsrisate, Sunee
Detection and differentiation of Burkholderia species with pathogenic potential in environmental soil samples
title Detection and differentiation of Burkholderia species with pathogenic potential in environmental soil samples
title_full Detection and differentiation of Burkholderia species with pathogenic potential in environmental soil samples
title_fullStr Detection and differentiation of Burkholderia species with pathogenic potential in environmental soil samples
title_full_unstemmed Detection and differentiation of Burkholderia species with pathogenic potential in environmental soil samples
title_short Detection and differentiation of Burkholderia species with pathogenic potential in environmental soil samples
title_sort detection and differentiation of burkholderia species with pathogenic potential in environmental soil samples
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7790303/
https://www.ncbi.nlm.nih.gov/pubmed/33411797
http://dx.doi.org/10.1371/journal.pone.0245175
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