Sensitive and specific molecular detection of Burkholderia pseudomallei, the causative agent of melioidosis, in the soil of tropical northern Australia
Sensitive and specific molecular detection of Burkholderia pseudomallei, the causative agent of melioidosis, in the soil of tropical northern Australia
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DOI：
10.1128/aem.01038-07
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发表时间：
2007-11-01
影响因子：
4.4
通讯作者：
Currie, Bart J.
中科院分区：
文献类型：
--
作者：
Kaestli, Mirjam;Mayo, Mark;Currie, Bart J.
类鼻疽伯克霍尔德氏菌（Burkholderia pseudomallei）是一种革兰氏阴性菌，生活在地方性地区如热带北方澳大利亚和东南亚的土壤和水中，是人类和动物中严重疾病类鼻疽的病因。感染主要通过接触潮湿的污染土壤发生。B的环境分布。在澳大利亚北方是否存在假鼻疽仍不清楚。我们开发并评估了直接土壤B。基于最近发表的靶向B的实时PCR的假鼻疽DNA检测方法。类鼻疽III型分泌系统。用B对不同土壤类型进行了测定。并通过比较B.从澳大利亚北方的达尔文农村地区随机采集的104份土壤样本的假鼻疽检出率与基于培养的检出率。我们发现直接土壤B。鼻疽假单胞菌DNA检测不仅比培养快得多，而且被证明更灵敏，没有明显的假阳性结果。该方法为检测B提供了新的工具。快速、高灵敏度和特异性地检测土壤样品中的类鼻疽，适用于大规模B。环境筛选研究或爆发情况下。此外，对收集的104个土壤样品的分析显示，B.假鼻疽阳性部位和动物在这些位置的存在，以及潮湿的红棕色至红灰色土壤。
Burkholderia pseudomallei, the cause of the severe disease melioidosis in humans and animals, is a gramnegative saprophyte living in soil and water of areas of endemicity such as tropical northern Australia and Southeast Asia. Infection occurs mainly by contact with wet contaminated soil. The environmental distribution of B. pseudomallei in northern Australia is still unclear. We developed and evaluated a direct soil B. pseudomallei DNA detection method based on the recently published real-time PCR targeting the B. pseudomallei type III secretion system. The method was evaluated by inoculating different soil types with B. pseudomallei dilution series and by comparing B. pseudomallei detection rate with culture-based detection rate for 104 randomly collected soil samples from the Darwin rural area in northern Australia. We found that direct soil B. pseudomallei DNA detection not only was substantially faster than culture but also proved to be more sensitive with no evident false-positive results. This assay provides a new tool to detect B. pseudomallei in soil samples in a fast and highly sensitive and specific manner and is applicable for large-scale B. pseudomallei environmental screening studies or in outbreak situations. Furthermore, analysis of the 104 collected soil samples revealed a significant association between B. pseudomallei-positive sites and the presence of animals at these locations and also with moist, reddish brown-to-reddish gray soils.