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一种用于鉴定六种医学上重要念珠菌属物种的单酶PCR-RFLP检测方法。

A one-enzyme PCR-RFLP assay for identification of six medically important Candida species.

作者信息

Mirhendi Hossein, Makimura Koichi, Khoramizadeh Mohamadreza, Yamaguchi Hideyo

机构信息

Department of Medical Parasitology and Mycology, School of Public Health and Institute of Public Health Researches (Esfahan Center), Tehran University of Medical Sciences, Tehran, Iran.

出版信息

Nihon Ishinkin Gakkai Zasshi. 2006;47(3):225-9. doi: 10.3314/jjmm.47.225.

Abstract

Early identification of Candida isolates to the species level is necessary for effective antifungal therapy, and can also facilitate control of hospital infections. Phenotype-based methods for identifying Candida species are often difficult and time-consuming. Molecular biological techniques provide a useful alternative approach. In the present study, the ITS1-5.8S-ITS2 regions of fungal rRNA genes were amplified with universal primers in 20 standard strains. Digestion of the PCR products with one restriction enzyme, MspI, allowed discrimination of medically important Candida species, including C. albicans, C. glabrata, C. parapsilosis, C. tropicalis, C. krusei, and C. guilliermondii. Using this method, we successfully identified 137 clinical isolates of Candida. Among them, C. albicans was identified as the most common species, followed by C. parapsilosis, C. tropicalis, C. glabrata, C. krusei, and C. guilliermondii. This method is a simple, rapid, and cost-effective method for differentiation between species that is applicable in clinical laboratories.

摘要

将念珠菌分离株鉴定到种水平对于有效的抗真菌治疗是必要的,并且还可以促进医院感染的控制。基于表型的念珠菌鉴定方法通常既困难又耗时。分子生物学技术提供了一种有用的替代方法。在本研究中,用通用引物在20株标准菌株中扩增真菌rRNA基因的ITS1-5.8S-ITS2区域。用一种限制性内切酶MspI消化PCR产物,可以区分医学上重要的念珠菌物种,包括白色念珠菌、光滑念珠菌、近平滑念珠菌、热带念珠菌、克柔念珠菌和季也蒙念珠菌。使用这种方法,我们成功鉴定了137株临床念珠菌分离株。其中,白色念珠菌被鉴定为最常见的物种,其次是近平滑念珠菌、热带念珠菌、光滑念珠菌、克柔念珠菌和季也蒙念珠菌。该方法是一种简单、快速且经济高效的区分物种的方法,适用于临床实验室。

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