Barbedo Leonardo Silva, Figueiredo-Carvalho Maria Helena Galdino, Muniz Mauro de Medeiros, Zancopé-Oliveira Rosely Maria
Laboratório de Micologia, Instituto Nacional de Infectologia Evandro Chagas, Fundação Oswaldo Cruz, Rio de Janeiro, RJ, Brasil.
Mem Inst Oswaldo Cruz. 2016 Apr;111(4):267-70. doi: 10.1590/0074-02760150466.
Currently, it is accepted that there are three species that were formerly grouped under Candida parapsilosis: C. para- psilosis sensu stricto, Candida orthopsilosis, and Candida metapsilosis. In fact, the antifungal susceptibility profiles and distinct virulence attributes demonstrate the differences in these nosocomial pathogens. An accurate, fast, and economical identification of fungal species has been the main goal in mycology. In the present study, we searched sequences that were available in the GenBank database in order to identify the complete sequence for the internal transcribed spacer (ITS)1-5.8S-ITS2 region, which is comprised of the forward and reverse primers ITS1 and ITS4. Subsequently, an in silico polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was performed to differentiate the C. parapsilosis complex species. Ninety-eight clinical isolates from patients with fungaemia were submitted for analysis, where 59 isolates were identified as C. parapsilosis sensu stricto, 37 were identified as C. orthopsilosis, and two were identified as C. metapsilosis. PCR-RFLP quickly and accurately identified C. parapsilosis complex species, making this method an alternative and routine identification system for use in clinical mycology laboratories.
目前，人们公认以前归在近平滑念珠菌名下的有三个菌种：狭义的近平滑念珠菌、正平滑念珠菌和间平滑念珠菌。事实上，这些医院感染病原体在抗真菌药敏谱和独特的毒力属性方面存在差异。准确、快速且经济地鉴定真菌菌种一直是真菌学的主要目标。在本研究中，我们在GenBank数据库中搜索可用序列，以确定由正向引物ITS1和反向引物ITS4组成的内转录间隔区（ITS）1-5.8S-ITS2区域的完整序列。随后，进行了虚拟聚合酶链反应-限制性片段长度多态性分析（PCR-RFLP），以区分近平滑念珠菌复合菌种。对98例真菌血症患者的临床分离株进行了分析，其中59株被鉴定为狭义的近平滑念珠菌，37株被鉴定为正平滑念珠菌，2株被鉴定为间平滑念珠菌。PCR-RFLP能快速准确地鉴定近平滑念珠菌复合菌种，使该方法成为临床真菌学实验室用于菌种鉴定的一种替代且常规的鉴定系统。
