Validation and clinical application of a molecular method for the identification of Cryptococcus neoformans/Cryptococcus gattii complex DNA in human clinical specimens

Vanessa Rivera, Marcela Gaviria, Cesar Muñoz-Cadavid, Luz Cano, Tonny Naranjo

    Research output: Contribution to journalArticle in an indexed scientific journalpeer-review

    13 Scopus citations

    Abstract

    The diagnosis of cryptococcosis is usually performed based on cultures of tissue or body fluids and isolation of the fungus, but this method may require several days. Direct microscopic examination, although rapid, is relatively insensitive. Biochemical and immunodiagnostic rapid tests are also used. However, all of these methods have limitations that may hinder final diagnosis. The increasing incidence of fungal infections has focused attention on tools for rapid and accurate diagnosis using molecular biological techniques. Currently, PCR-based methods, particularly nested, multiplex and real-time PCR, provide both high sensitivity and specificity.In the present study, we evaluated a nested PCR targeting the gene encoding the ITS-1 and ITS-2 regions of rDNA in samples from a cohort of patients diagnosed with cryptococcosis. The results showed that in our hands, this Cryptococcus nested PCR assay has 100% specificity and 100% sensitivity and was able to detect until 2 femtograms of Cryptococcus DNA.

    Original languageEnglish
    Pages (from-to)563-570
    Number of pages8
    JournalBrazilian Journal of Infectious Diseases
    Volume19
    Issue number6
    DOIs
    StatePublished - 1 Nov 2015

    Bibliographical note

    Publisher Copyright:
    © 2015 Elsevier Editora Ltda.

    Keywords

    • Cryptococcosis
    • Cryptococcus neoformans/Cryptococcus gattii complex
    • Molecular diagnosis
    • Nested PCR

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