de Andrade Tânia Sueli, Cury Arlete Emily, de Castro Luiz Guilherme Martins, Hirata Mario Hiroyuki, Hirata Rosario Dominguez Crespo
Departamento de Análises Clínicas e Toxicológicas, Faculdade de Ciências Farmacêuticas, Universidade de São Paulo, São Paulo, Brasil.
Diagn Microbiol Infect Dis. 2007 Mar;57(3):267-72. doi: 10.1016/j.diagmicrobio.2006.08.024.
Fonsecaea pedrosoi is the most common etiologic agent of chromoblastomycosis. F. pedrosoi and other dematiaceous fungi are usually identified by morphologic studies. We have developed a duplex polymerase chain reaction (PCR) targeting the ribosomal DNA for rapid and more specific identification of the genus Fonsecaea. DNA samples from 103 isolates of Fonsecaea species and other dematiaceous fungi were amplified by PCR using universal and specific primers targeting ITS1-5.8S-ITS2 region of the ribosomal DNA. Universal primers were used for detection of non-Fonsecaea DNA. Fonsecaea-specific PCR product was found in 70 (68.0%) isolates including 4 strains that did not develop conidiogenesis. Thirty non-Fonsecaea and 3 Fonsecaea compacta isolates were negative by duplex PCR. These results were confirmed by DNA sequencing analysis indicating the high specificity of the duplex PCR assay. In conclusion, the duplex PCR is a rapid and specific assay for identification of Fonsecaea isolates mainly for the strains that are difficult to identify by morphologic methods.
裴氏着色真菌是着色芽生菌病最常见的病原体。裴氏着色真菌和其他暗色真菌通常通过形态学研究来鉴定。我们开发了一种针对核糖体DNA的双重聚合酶链反应（PCR），用于快速且更特异性地鉴定裴氏着色真菌属。使用针对核糖体DNA的ITS1-5.8S-ITS2区域的通用引物和特异性引物，通过PCR对来自103株裴氏着色真菌属菌株和其他暗色真菌的DNA样本进行扩增。通用引物用于检测非裴氏着色真菌的DNA。在70株（68.0%）菌株中发现了裴氏着色真菌特异性PCR产物，其中包括4株未产生分生孢子形成的菌株。30株非裴氏着色真菌和3株紧密裴氏着色真菌菌株通过双重PCR检测为阴性。DNA测序分析证实了这些结果，表明双重PCR检测具有高度特异性。总之，双重PCR是一种快速且特异性的检测方法，主要用于鉴定难以通过形态学方法鉴定的裴氏着色真菌菌株。
