Rapid identification of Fonsecaea by duplex polymerase chain reaction in isolates from patients with chromoblastomycosis

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.