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随机扩增多态性DNA(RAPD)及限制性内切酶分析在白色念珠菌口腔携带研究中的应用

Application of RAPD and restriction enzyme analysis to the study of oral carriage of Candida albicans.

作者信息

Howell S A, Anthony R M, Power E

机构信息

Department of Microbial Diseases, St John's Institute of Dermatology, London, UK.

出版信息

Lett Appl Microbiol. 1996 Feb;22(2):125-8. doi: 10.1111/j.1472-765x.1996.tb01124.x.

Abstract

The genetic similarity of nineteen isolates of Candida albicans from four patients were compared by restriction fragment length polymorphism (RFLP) using EcoRI or HinfI, which both detected five types, and by random amplification of polymorphic DNA (RAPD), which detected three types. Phenotypically unusual isolates also produced distinct patterns with both typing systems demonstrating the carriage of two groups of C. albicans as well as the presence of more than one type in some subjects. Methods of DNA preparation were compared for the production of reproducible patterns; including using the supernatant fluid of boiled intact or spheroplasted cells for RAPD, and DNA precipitated from chloroform extracted cell lysate for RFLP and RAPD. Consistent patterns were produced from the DNA precipitate by RAPD and after an additional precipitation by RFLP, thus removing the necessity for lengthy extraction procedures or the use of toxic chemicals for purification.

摘要

使用EcoRI或HinfI通过限制性片段长度多态性(RFLP)比较了来自四名患者的19株白色念珠菌分离株的遗传相似性,二者均检测到五种类型,同时通过随机扩增多态性DNA(RAPD)进行比较,后者检测到三种类型。表型异常的分离株在两种分型系统中也产生了不同的模式,表明携带两组白色念珠菌以及某些受试者中存在不止一种类型。比较了DNA制备方法以产生可重复的模式;包括使用煮沸的完整细胞或原生质球的上清液进行RAPD,以及从氯仿提取的细胞裂解物中沉淀的DNA用于RFLP和RAPD。通过RAPD从DNA沉淀物中产生一致的模式,通过RFLP进行额外沉淀后也产生一致的模式,从而无需冗长的提取程序或使用有毒化学物质进行纯化。

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