Direct detection of Burkholderia pseudomallei and biological factors in soil

Rasana W. Sermswan, Phairat Royros, Nittaya Khakhum, Surasakdi Wongratanacheewin, Apichai Tuanyok

Research output: Contribution to journalArticlepeer-review

16 Scopus citations

Abstract

Background: Burkholderia pseudomallei, a Gram-negative saprophytic bacillus, is a severe infectious agent that causes melioidosis and soil is the most important reservoir. Methods: One hundred and forty soil samples were tested for pH, moisture content and total C and N measurements and used for DNA extraction and culture for B. pseudomallei. The quantitative real-time PCR (qPCR) targeting wcbG, a putative capsular polysaccharide biosynthesis protein gene of B. pseudomallei, was developed to detect the bacterium, and random amplified polymorphic DNA (RAPD) was used to detect the microbial diversity in soil. Results: The acidic pH was correlated with the presence of the bacterium. Forty-four soil sites (44/140, 31.4%) were positive for B. pseudomallei by qPCR, of which 21 were positive by culture. The limit of detection is 32 fg of DNA (about 4 genomes). The RAPD method could classify the soil samples into low diversity (LD) and high diversity (HD) sites. The trend of LD was found with B. pseudomallei positive soil sites. Conclusions: The acidity of the soil or metabolites from organisms in the sites may contribute to the presence of the bacterium. Further investigation of microbes by a more robust method should elucidate biological factors that promote the presence of B. pseudomallei and may be used for controlling the bacterium in the environment.

Original languageEnglish (US)
Pages (from-to)462-468
Number of pages7
JournalTransactions of the Royal Society of Tropical Medicine and Hygiene
Volume109
Issue number7
DOIs
StatePublished - May 18 2015
Externally publishedYes

Keywords

  • Burkholderia pseudomallei
  • Diversity
  • Quantitative real-time PCR
  • Random amplified polymorphic DNA
  • Soil

ASJC Scopus subject areas

  • Parasitology
  • Public Health, Environmental and Occupational Health
  • Infectious Diseases

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