摘要
介绍了一种基于聚合酶链反应(PCR)结合限制性内切酶分析技术区分临床重要真菌的方法。首先利用9种真菌的通用引物扩增其18SrRNA基因,然后用HaeⅢ、BglⅠ和NheⅠ三种限制性内切酶将扩增产物消化成不同数量和不同大小的DNA片段,可以对白色念珠菌、新型隐球菌和烟曲霉菌等9种临床上常见的医学真菌作出快速鉴定。该技术具有快速、敏感、不需杂交、可在24h以内完成鉴定等优点,有良好的临床应用前景。
The methods of identification of medically important fungi to the species level currently used are examination of morphology or determination of biochemical characteristics. Both methods require the presence of viable fungi and are insensitive. In this paper,a rapid method for identifying 9 species of medically important fungi (Candida albicans, C. tropicalis, C. kefyr, C. parapsilosis, Torulopsis glabrata, C. lipolytica, Cryptococcus neoformans, Aspergillus fumigatus, A. flavus) using the polymerase chain reaction (PCR) and restriction enzyme analysis is described. The primers we designed amplified the 621-687 bp fragments of the 18 S rRNA gene of 9 fungal species. The PCR products were then digested by HaeⅢ, BglⅠ and NheⅠ. The 9 species of fungi mentioned above can be differentiated to the species level. It is a rapid and sensitive method, which does not involve hybridization steps or radioactivity, and the identification can be completed within one day.
出处
《军事医学科学院院刊》
CSCD
北大核心
1997年第1期54-57,共4页
Bulletin of the Academy of Military Medical Sciences