Livério Harisson Oliveira, Ruiz Luciana da Silva, Freitas Roseli Santos de, Nishikaku Angela, Souza Ana Clara de, Paula Claudete Rodrigues, Domaneschi Carina
Universidade de São Paulo, Faculdade de Odontologia, Departamento de Estomatologia, São Paulo, São Paulo, Brazil.
Instituto Adolfo Lutz, Núcleo de Ciências Biomédicas, Bauru, São Paulo, Brazil.
Rev Inst Med Trop Sao Paulo. 2017 Apr 13;59:e14. doi: 10.1590/S1678-9946201759014.
The aim of this study was to assess a collection of yeasts to verify the presence of Candida dubliniensis among strains isolated from the oral mucosa of AIDS pediatric patients which were initially characterized as Candida albicans by the traditional phenotypic method, as well as to evaluate the main phenotypic methods used in the discrimination between the two species and confirm the identification through genotypic techniques, i.e., DNA sequencing. Twenty-nine samples of C. albicans isolated from this population and kept in a fungi collection were evaluated and re-characterized. In order to differentiate the two species, phenotypic tests (Thermotolerance tests, Chromogenic medium, Staib agar, Tobacco agar, Hypertonic medium) were performed and genotypic techniques using DNA sequencing were employed for confirmation of isolated species. Susceptibility and specificity were calculated for each test. No phenotypic test alone was sufficient to provide definitive identification of C. dubliniensis or C. albicans, as opposed to results of molecular tests. After amplification and sequencing of specific regions of the 29 studied strains, 93.1% of the isolates were identified as C. albicans and 6.9% as C. dubliniensis. The Staib agar assay showed a higher susceptibility (96.3%) in comparison with other phenotypic techniques. Therefore, genotypic methods are indispensable for the conclusive identification and differentiation between these species.
本研究的目的是评估一组酵母,以验证从艾滋病儿童患者口腔黏膜分离出的菌株中是否存在都柏林念珠菌,这些菌株最初通过传统表型方法鉴定为白色念珠菌,同时评估用于区分这两种念珠菌的主要表型方法,并通过基因分型技术(即DNA测序)确认鉴定结果。对从该人群中分离并保存在真菌收藏中的29株白色念珠菌样本进行了评估和重新鉴定。为了区分这两个菌种,进行了表型试验(耐热性试验、显色培养基、施泰布琼脂、烟草琼脂、高渗培养基),并采用DNA测序的基因分型技术对分离出的菌种进行确认。计算了每个试验的敏感性和特异性。与分子试验结果相反,没有任何一种表型试验足以明确鉴定都柏林念珠菌或白色念珠菌。对29株研究菌株的特定区域进行扩增和测序后,93.1%的分离株被鉴定为白色念珠菌,6.9%为都柏林念珠菌。与其他表型技术相比,施泰布琼脂试验显示出更高的敏感性(96.3%)。因此,基因分型方法对于这些菌种的确切鉴定和区分是必不可少的。