Kilvington S, Beeching J
Public Health Laboratory, Royal United Hospital, Bath, England.
Appl Environ Microbiol. 1995 Jun;61(6):2071-8. doi: 10.1128/aem.61.6.2071-2078.1995.
Naegleria fowleri is a small free-living amoeboflagellate found in warm water habitats worldwide. The organism is pathogenic to humans, causing fatal primary amoebic meningoencephalitis. When monitoring the environment for the presence of N. fowleri, it is important to reliably differentiate the organism from other closely related but nonpathogenic species. To this end, we have developed species-specific DNA probes for use in the rapid identification of N. fowleri from the environment. Samples were taken from the thermal springs in Bath, England, and cultured for amoebae. Of 84 isolates of thermophilic Naegleria spp., 10 were identified as N. fowleri by probe hybridization. The identity of these isolates was subsequently confirmed by their specific whole-cell DNA restriction fragment length polymorphisms (RFLPs). One DNA clone was found to contain a repeated element that detected chromosomal RFLPs that were not directly visible on agarose gels. This enabled the further differentiation of strains within geographically defined whole-cell DNA RFLP groups. N. fowleri DNA probes represent a specific and potentially rapid method for the identification of the organism soon after primary isolation from the environment.
福氏耐格里阿米巴是一种小型自由生活的变形鞭毛虫,在世界各地的温水生境中均有发现。该生物体对人类具有致病性,可导致致命的原发性阿米巴脑膜脑炎。在监测环境中是否存在福氏耐格里阿米巴时,可靠地区分该生物体与其他密切相关但无致病性的物种非常重要。为此,我们开发了物种特异性DNA探针,用于从环境中快速鉴定福氏耐格里阿米巴。样本取自英国巴斯的温泉,并培养其中的阿米巴。在84株嗜热耐格里属物种的分离株中,通过探针杂交鉴定出10株为福氏耐格里阿米巴。这些分离株的身份随后通过其特定的全细胞DNA限制性片段长度多态性(RFLP)得以确认。发现一个DNA克隆含有一个重复元件,该元件可检测到在琼脂糖凝胶上无法直接看到的染色体RFLP。这使得能够在地理定义的全细胞DNA RFLP组内进一步区分菌株。福氏耐格里阿米巴DNA探针是一种在从环境中初次分离后不久就可用于鉴定该生物体的特异性且可能快速的方法。