Elwin K, Chalmers R M, Roberts R, Guy E C, Casemore D P
PHLS Cryptosporidium Reference Unit, Swansea Public Health Laboratory, Singleton Hospital, Swansea, United Kingdom.
Appl Environ Microbiol. 2001 Dec;67(12):5581-4. doi: 10.1128/AEM.67.12.5581-5584.2001.
The application of genotyping to clinical isolates of Cryptosporidium has increased significantly our knowledge and understanding of the distribution and epidemiology of this parasite. However, some methods can be laborious and demand specialist technical expertise. PCR-restriction fragment length polymorphism (RFLP) techniques represent a more rapid and simple method of genotyping to support epidemiological and clinical investigations than conventional DNA analytical techniques. We describe a nested PCR-RFLP technique that identifies polymorphisms in the C. parvum thrombospondin-related adhesive protein gene locus; this method offers a sensitive and specific tool for the confirmation and investigation of disease associated with C. parvum. The potential of this enhanced method is demonstrated by its application to the confirmation and epidemiological investigation of an outbreak of cryptosporidiosis associated with a school visit to an open farm.
基因分型技术在隐孢子虫临床分离株中的应用，显著增加了我们对这种寄生虫的分布和流行病学的认识与理解。然而，一些方法可能很繁琐，且需要专业技术专长。与传统的DNA分析技术相比，聚合酶链反应-限制性片段长度多态性（PCR-RFLP）技术是一种更快速、简单的基因分型方法，可支持流行病学和临床研究。我们描述了一种巢式PCR-RFLP技术，该技术可识别微小隐孢子虫血小板反应蛋白相关黏附蛋白基因位点的多态性；此方法为与微小隐孢子虫相关疾病的确诊和调查提供了一种灵敏且特异的工具。通过将这种改进方法应用于一次与学校参观开放式农场相关的隐孢子虫病暴发的确诊和流行病学调查，证明了其潜力。
