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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th-mahidol.794162022-08-04T18:42:13Z Detection and differentiation of Burkholderia species with pathogenic potential in environmental soil samples Sujintana Janesomboon Veerachat Muangsombut Varintip Srinon Chatruthai Meethai Chayada S. Tharinjaroen Premjit Amornchai Patoo Withatanung Narisara Chantratita Mark Mayo Vanaporn Wuthiekanun Bart J. Currie Joanne M. Stevens Sunee Korbsrisate Siriraj Hospital University of Edinburgh, Roslin Institute Menzies School of Health Research Mahidol University Royal Darwin Hospital Chiang Mai University Multidisciplinary 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. 2022-08-04T11:42:13Z 2022-08-04T11:42:13Z 2021-01-01 Article PLoS ONE. Vol.16, No.1 January (2021) 10.1371/journal.pone.0245175 19326203 2-s2.0-85099351792 https://repository.li.mahidol.ac.th/handle/123456789/79416 Mahidol University SCOPUS https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85099351792&origin=inward |
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Multidisciplinary Sujintana Janesomboon Veerachat Muangsombut Varintip Srinon Chatruthai Meethai Chayada S. Tharinjaroen Premjit Amornchai Patoo Withatanung Narisara Chantratita Mark Mayo Vanaporn Wuthiekanun Bart J. Currie Joanne M. Stevens Sunee Korbsrisate Detection and differentiation of Burkholderia species with pathogenic potential in environmental soil samples |
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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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Siriraj Hospital |
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Siriraj Hospital Sujintana Janesomboon Veerachat Muangsombut Varintip Srinon Chatruthai Meethai Chayada S. Tharinjaroen Premjit Amornchai Patoo Withatanung Narisara Chantratita Mark Mayo Vanaporn Wuthiekanun Bart J. Currie Joanne M. Stevens Sunee Korbsrisate |
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Article |
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Sujintana Janesomboon Veerachat Muangsombut Varintip Srinon Chatruthai Meethai Chayada S. Tharinjaroen Premjit Amornchai Patoo Withatanung Narisara Chantratita Mark Mayo Vanaporn Wuthiekanun Bart J. Currie Joanne M. Stevens Sunee Korbsrisate |
author_sort |
Sujintana Janesomboon |
title |
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_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_sort |
detection and differentiation of burkholderia species with pathogenic potential in environmental soil samples |
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2022 |
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https://repository.li.mahidol.ac.th/handle/123456789/79416 |
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1763490299627175936 |