title: Molecular methods for the diagnosis and characterization of Cryptococcus: a review.
abstract: Cryptococcosis is a fungal infection caused by yeasts of the genus Cryptococcus, with Cryptococcus neoformans and Cryptococcus gattii as the primary pathogenic species. This disease is a threat to immunocompromised patients, especially those who have AIDS. However, the disease has also been described in healthy individuals. The tests used to identify these microorganisms have limitations that make final diagnosis difficult. However, currently there are specific gene sequences that can be used to detect C. neoformans and C. gattii from clinical specimens and cultures. These sequences can be used for identification, typing, and the study of population genetics. Among the main identification techniques are hybridization, which was the pioneer in molecular identification and development of specific probes for pathogen detection; PCR and other PCR-based methods, particularly nested PCR and multiplex PCR; and sequencing of specific genomic regions that are amplified through PCR, which is especially useful for diagnosis of cryptococcosis caused by unconventional Cryptococcus sp. Concerning microorganism typing, the following techniques have shown the best ability to differentiate between fungal serotypes and molecular types: PCR fingerprinting, PCR-RFLP, AFLP, and MLST. Thus, the accumulation of data generated by molecular methods can have a positive impact on monitoring resistant strains and treating diseases.
authors.affiliation: Specialized Medical Mycology Center, Federal University of Ceará, Rua Coronel Nunes de Melo, Rodolfo Teófilo, Fortaleza, Ceará, Brazil.
authors.forename: José Júlio Costa
authors.initials: JJ
authors.lastname: Sidrim
authors.forename: Ana Karoline Freire
authors.initials: AK
authors.lastname: Costa
authors.forename: Rossana Aguiar
authors.initials: RA
authors.lastname: Cordeiro
authors.forename: Raimunda Sâmia Nogueira
authors.initials: RS
authors.lastname: Brilhante
authors.forename: Fernanda Edna Araújo
authors.initials: FE
authors.lastname: Moura
authors.forename: Débora Souza Collares Maia
authors.initials: DS
authors.lastname: Castelo-Branco
authors.forename: Manoel Paiva de Araújo
authors.initials: MP
authors.lastname: Neto
authors.forename: Marcos Fábio Gadelha
authors.initials: MF
authors.lastname: Rocha
mesh_terms: D003453:Cryptococcosis / Q000175:diagnosis*; D003454:Cryptococcus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D016172:DNA Fingerprinting / Q000379:methods; D004271:DNA, Fungal / Q000235:genetics; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D016533:Mycological Typing Techniques; D009172:Mycology / Q000379:methods*
chemical_list: D004271:DNA, Fungal