Sermswan Rasana W, Royros Phairat, Khakhum Nittaya, Wongratanacheewin Surasakdi, Tuanyok Apichai
Department of Biochemistry, Faculty of Medicine, Khon Kean University, 123 Mitraparp Rd, Khon Kaen 40002, Thailand Melioidosis Research Center, Faculty of Medicine, Khon Kean University, Khon Kaen, Thailand
Department of Biochemistry, Faculty of Medicine, Khon Kean University, 123 Mitraparp Rd, Khon Kaen 40002, Thailand Melioidosis Research Center, Faculty of Medicine, Khon Kean University, Khon Kaen, Thailand.
Trans R Soc Trop Med Hyg. 2015 Jul;109(7):462-8. doi: 10.1093/trstmh/trv040. Epub 2015 Jun 4.
Burkholderia pseudomallei, a Gram-negative saprophytic bacillus, is a severe infectious agent that causes melioidosis and soil is the most important reservoir.
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.
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.
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.
类鼻疽伯克霍尔德菌是一种革兰氏阴性腐生杆菌，是引起类鼻疽的严重感染病原体，土壤是其最重要的储存宿主。
对140份土壤样本进行pH值、水分含量以及总碳和总氮测量，并用于提取DNA以及培养类鼻疽伯克霍尔德菌。开发了针对类鼻疽伯克霍尔德菌假定的荚膜多糖生物合成蛋白基因wcbG的定量实时PCR（qPCR）来检测该细菌，并用随机扩增多态性DNA（RAPD）检测土壤中的微生物多样性。
酸性pH值与该细菌的存在相关。通过qPCR检测，44个土壤位点（44/140，31.4%）类鼻疽伯克霍尔德菌呈阳性，其中21个通过培养呈阳性。检测限为32 fg DNA（约4个基因组）。RAPD方法可将土壤样本分为低多样性（LD）和高多样性（HD）位点。在类鼻疽伯克霍尔德菌阳性土壤位点发现了LD趋势。
土壤的酸度或位点中生物体的代谢产物可能导致该细菌的存在。通过更强大的方法对微生物进行进一步研究应能阐明促进类鼻疽伯克霍尔德菌存在的生物学因素，并可用于控制环境中的该细菌。
