van Belkum A, De Jonckheere J, Quint W G
Department of Molecular Biology, Diagnostic Centre SSDZ, Delft, The Netherlands.
J Clin Microbiol. 1992 Oct;30(10):2595-8. doi: 10.1128/jcm.30.10.2595-2598.1992.
All six Naegleria species recognized to date were studied by interrepeat polymerase chain reaction (PCR). Priming at repeat sequences, which are known to be variable among eukaryotes, yielded electrophoretic DNA banding patterns that were specific for any single species. With a single PCR and simple gel electrophoresis, species determination could be performed in less than 1 day. Unambiguous discrimination between the pathogen N. fowleri and nonpathogenic Naegleria species appeared to be possible. Analysis of DNAs obtained from 20 separate isolates of N. fowleri revealed that geographic variation of the genetic fingerprints rarely occurs. All but 3 of 20 isolates of N. fowleri which were investigated showed identical banding patterns; for two isolates from New Zealand and one from Australia, a limited number of additional bands was detected, independent of the PCR primers used. These data corroborate previous findings on the genetic stability of pathogenic N. fowleri.
利用重复序列间聚合酶链反应(PCR)对迄今已识别出的所有六种耐格里属物种进行了研究。在真核生物中已知具有变异性的重复序列处进行引物设计,产生了对任何单一物种具有特异性的电泳DNA条带模式。通过一次PCR和简单的凝胶电泳,可在不到一天的时间内完成物种鉴定。似乎有可能明确区分致病性福氏耐格里阿米巴与非致病性耐格里属物种。对从20个独立的福氏耐格里阿米巴分离株获得的DNA进行分析发现,遗传指纹的地理变异很少发生。在20个被研究的福氏耐格里阿米巴分离株中,除3个外,其余所有分离株均显示出相同的条带模式;对于来自新西兰的两个分离株和来自澳大利亚的一个分离株,无论使用何种PCR引物,均检测到数量有限的额外条带。这些数据证实了先前关于致病性福氏耐格里阿米巴遗传稳定性的研究结果。