{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,4]],"date-time":"2026-10-04T05:12:49Z","timestamp":1791090769956,"version":"4.1.0"},"reference-count":63,"publisher":"American Society for Microbiology","issue":"18","license":[{"start":{"date-parts":[[2011,9,15]],"date-time":"2011-09-15T00:00:00Z","timestamp":1316044800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Appl Environ Microbiol"],"published-print":{"date-parts":[[2011,9,15]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            The soil bacterium and potential biothreat agent\n            <jats:named-content content-type=\"genus-species\">Burkholderia pseudomallei<\/jats:named-content>\n            causes the infectious disease melioidosis, which is naturally acquired through environmental contact with the bacterium. Environmental detection of\n            <jats:named-content content-type=\"genus-species\">B. pseudomallei<\/jats:named-content>\n            represents the basis for the development of a geographical risk map for humans and livestock. The aim of the present study was to develop a highly sensitive, culture-independent, DNA-based method that allows direct quantification of\n            <jats:named-content content-type=\"genus-species\">B. pseudomallei<\/jats:named-content>\n            from soil. We established a protocol for\n            <jats:named-content content-type=\"genus-species\">B. pseudomallei<\/jats:named-content>\n            soil DNA isolation, purification, and quantification by quantitative PCR (qPCR) targeting a type three secretion system 1 single-copy gene. This assay was validated using 40 soil samples from Northeast Thailand that underwent parallel bacteriological culture. All 26 samples that were\n            <jats:named-content content-type=\"genus-species\">B. pseudomallei<\/jats:named-content>\n            positive by direct culture were\n            <jats:named-content content-type=\"genus-species\">B. pseudomallei<\/jats:named-content>\n            qPCR positive, with a median of 1.84 \u00d7 10\n            <jats:sup>4<\/jats:sup>\n            genome equivalents (range, 3.65 \u00d7 10\n            <jats:sup>2<\/jats:sup>\n            to 7.85 \u00d7 10\n            <jats:sup>5<\/jats:sup>\n            ) per gram of soil, assuming complete recovery of DNA. This was 10.6-fold (geometric mean; range, 1.1- to 151.3-fold) higher than the bacterial count defined by direct culture. Moreover, the qPCR detected\n            <jats:named-content content-type=\"genus-species\">B. pseudomallei<\/jats:named-content>\n            in seven samples (median, 36.9 genome equivalents per g of soil; range, 9.4 to 47.3) which were negative by direct culture. These seven positive results were reproduced using a nested PCR targeting a second, independent\n            <jats:named-content content-type=\"genus-species\">B. pseudomallei<\/jats:named-content>\n            -specific sequence. Two samples were direct culture and qPCR negative but nested PCR positive. Five samples were negative by both PCR methods and culture. In conclusion, our PCR-based system provides a highly specific and sensitive tool for the quantitative environmental surveillance of\n            <jats:named-content content-type=\"genus-species\">B. pseudomallei<\/jats:named-content>\n            .\n          <\/jats:p>","DOI":"10.1128\/aem.00735-11","type":"journal-article","created":{"date-parts":[[2011,7,30]],"date-time":"2011-07-30T04:16:14Z","timestamp":1311999374000},"page":"6486-6494","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":53,"title":["Highly Sensitive Direct Detection and Quantification of Burkholderia pseudomallei Bacteria in Environmental Soil Samples by Using Real-Time PCR"],"prefix":"10.1128","volume":"77","author":[{"given":"Trinh Thanh","family":"Trung","sequence":"first","affiliation":[{"name":"Friedrich Loeffler Institute of Medical Microbiology, Ernst Moritz Arndt University of Greifswald, Greifswald, 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