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Regarded as the only pathogenic members of this group, <jats:italic>B<\/jats:italic>. <jats:italic>pseudomallei<\/jats:italic> and <jats:italic>B<\/jats:italic>. <jats:italic>mallei<\/jats:italic> cause the diseases melioidosis and glanders, respectively. Additionally, variant strains of <jats:italic>B<\/jats:italic>. <jats:italic>pseudomallei<\/jats:italic> and <jats:italic>B<\/jats:italic>. <jats:italic>thailandensis<\/jats:italic> exist that include the geographically restricted <jats:italic>B<\/jats:italic>. <jats:italic>pseudomallei<\/jats:italic> that express a <jats:italic>B<\/jats:italic>. <jats:italic>mallei<\/jats:italic>-like BimA protein (BPBM), and <jats:italic>B<\/jats:italic>. <jats:italic>thailandensis<\/jats:italic> that express a <jats:italic>B<\/jats:italic>. <jats:italic>pseudomallei<\/jats:italic>-like capsular polysaccharide (BTCV). To establish a PCR-based assay for the detection of pathogenic <jats:italic>Burkholderia<\/jats:italic> species or their variants, five PCR primers were designed to amplify species-specific sequences within the <jats:italic>bimA<\/jats:italic> (<jats:underline><jats:italic>B<\/jats:italic><\/jats:underline><jats:italic>urkholderia<\/jats:italic> <jats:underline>i<\/jats:underline>ntracellular <jats:underline>m<\/jats:underline>otility <jats:underline>A<\/jats:underline>) gene. Our multiplex PCR assay could distinguish pathogenic <jats:italic>B<\/jats:italic>. <jats:italic>pseudomallei<\/jats:italic> and BPBM from the non-pathogenic <jats:italic>B<\/jats:italic>. <jats:italic>thailandensis<\/jats:italic> and the BTCV strains. A second singleplex PCR successfully discriminated the BTCV from <jats:italic>B<\/jats:italic>. <jats:italic>thailandensis<\/jats:italic>. Apart from <jats:italic>B<\/jats:italic>. <jats:italic>humptydooensis<\/jats:italic>, specificity testing against other <jats:italic>Burkholderia<\/jats:italic> 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 <jats:italic>B<\/jats:italic>. <jats:italic>pseudomallei<\/jats:italic> and <jats:italic>B<\/jats:italic>. <jats:italic>thailandensis<\/jats:italic> were 5 and 6 CFU\/g soil, respectively. Furthermore, comparison between standard bacterial culture and the multiplex PCR to detect <jats:italic>B<\/jats:italic>. <jats:italic>pseudomallei<\/jats:italic> 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 <jats:italic>B<\/jats:italic>. <jats:italic>pseudomallei<\/jats:italic> and closely related members with pathogenic potential in soil.<\/jats:p>","DOI":"10.1371\/journal.pone.0245175","type":"journal-article","created":{"date-parts":[[2021,1,7]],"date-time":"2021-01-07T21:19:55Z","timestamp":1610054395000},"page":"e0245175","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":8,"title":["Detection and differentiation of Burkholderia species with pathogenic potential in environmental soil 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