Phenotypic and Molecular Methods for the Identification of Candida Dubliniensis
Keywords:
Candida dubliniensis; oral candidiasis; HIV/AIDS; polymerase chain reaction, phenotypic identification.Abstract
Introduction: Species of the genus Candida can cause a wide spectrum of clinical manifestations; although Candida albicans remain the most significant, other species are being isolated with increasing frequency. Among the latter, Candida dubliniensis is known to be closely phylogenetically related to Candida albicans and can develop resistance to azoles, making its correct identification essential. While various phenotypic methods have been used, molecular techniques define its diagnosis.
Objectives: To identify clinical isolates of the Candida albicans complex from the pathogenic fungi collection at the National Mycology Reference Laboratory of Pedro Kourí Institute of Tropical Medicine, and to establish a workflow algorithm to differentiate Candida albicans from Candida dubliniensis.
Methods: Forty-five isolates previously identified as Candida albicans, obtained from oral lesions of patients with HIV/AIDS, were re-identified. Ten phenotypic methods were evaluated, and polymerase chain reaction was applied to identify the rRNA ITS1 and ITS2 genes and the gene encoding Candida topoisomerase II.
Results: Filamentation and chlamydospore production, growth on tobacco agar with Tween 80, enzymatic activity, and growth on Sabouraud triphenyltetrazolium agar allowed for the presumptive identification of four Candida dubliniensis isolates; this was confirmed using molecular technique. Based on these tools, a laboratory algorithm was established to differentiate between the two species.
Conclusions: These findings confirm the need to combine phenotypic and genotypic methods to achieve conclusive differentiation between Candida albicans and Candida dubliniensis.
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