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通过随机扩增多态性DNA和核糖体DNA重复序列的限制性片段长度多态性分析对念珠菌属进行分子分型。

Molecular typing of Candida spp. by random amplification of polymorphic DNA and analysis of restriction fragment length polymorphism of ribosomal DNA repeats.

作者信息

Cresti S, Posteraro B, Sanguinetti M, Guglielmetti P, Rossolini G M, Morace G, Fadda G

机构信息

Dipartimento di Biologia Molecolare, Università di Siena, Italy.

出版信息

New Microbiol. 1999 Jan;22(1):41-52.

Abstract

Random amplification of polymorphic DNA (RAPD) using an arbitrary oligonucleotide primer (5'-CGGTGCGACG) and analysis of restriction fragment length polymorphism of ribosomal DNA (rDNA-RFLP) after digestion of genomic DNA with restriction endonuclease EcoRI were investigated as tools for genotypic delineation beyond the species level of 91 Candida clinical isolates and four reference strains including 33 Candida albicans, 19 Candida tropicalis, 22 Candida krusei and 21 Candida (Torulopsis) glabrata. Results indicated that both techniques can be useful for typing isolates of the above species, although showing a variable discriminative potential with different species. As compared to RAPD fingerprinting, the discriminative potential of rDNA-RFLP appeared to be highest for C. albicans and lowest for C. glabrata, being overall similar for C. krusei and identical for C. tropicalis. A comparative analysis of the results obtained with the two typing techniques showed that, except for C. tropicalis, they were able to provide non-redundant information, and that their use in combination could enhance the discriminative potential for delineation among C. glabrata and C. krusei isolates.

摘要

研究了使用任意寡核苷酸引物(5'-CGGTGCGACG)进行随机扩增多态性DNA(RAPD)以及用限制性内切酶EcoRI消化基因组DNA后分析核糖体DNA的限制性片段长度多态性(rDNA-RFLP),以此作为对91株念珠菌临床分离株和4株参考菌株进行种水平以上基因型鉴定的工具,这些菌株包括33株白色念珠菌、19株热带念珠菌、22株克鲁斯念珠菌和21株光滑念珠菌(假丝酵母属)。结果表明,这两种技术都可用于上述菌种分离株的分型,尽管对不同菌种显示出不同的鉴别潜力。与RAPD指纹图谱相比,rDNA-RFLP的鉴别潜力对白色念珠菌似乎最高,对光滑念珠菌最低,对克鲁斯念珠菌总体相似,对热带念珠菌则相同。对这两种分型技术所得结果的比较分析表明,除热带念珠菌外,它们能够提供非冗余信息,并且联合使用它们可以提高对光滑念珠菌和克鲁斯念珠菌分离株进行鉴别分型的潜力。

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