title: Validation and clinical application of a molecular method for the identification of Cryptococcus neoformans/Cryptococcus gattii complex DNA in human clinical specimens.
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.
authors.affiliation: Grupo de Micología Médica y Experimental, Corporación para Investigaciones Biológicas, Medellín, Colombia.
authors.forename: Vanessa
authors.initials: V
authors.lastname: Rivera
authors.affiliation: Grupo de Micología Médica y Experimental, Corporación para Investigaciones Biológicas, Medellín, Colombia.
authors.forename: Marcela
authors.initials: M
authors.lastname: Gaviria
authors.affiliation: Grupo de Micología Médica y Experimental, Corporación para Investigaciones Biológicas, Medellín, Colombia.
authors.forename: Cesar
authors.initials: C
authors.lastname: Muñoz-Cadavid
authors.affiliation: Grupo de Micología Médica y Experimental, Corporación para Investigaciones Biológicas, Medellín, Colombia; Escuela de Microbiología, Universidad de Antioquia, Medellín, Colombia.
authors.forename: Luz
authors.initials: L
authors.lastname: Cano
authors.affiliation: Grupo de Micología Médica y Experimental, Corporación para Investigaciones Biológicas, Medellín, Colombia; Escuela de Ciencias de la Salud, Universidad Pontifica Bolivariana, Medellín, Colombia. Electronic address: tnaranjo@cib.org.co.
authors.forename: Tonny
authors.initials: T
authors.lastname: Naranjo
mesh_terms: D003453:Cryptococcosis / Q000175:diagnosis* / Q000382:microbiology; D056285:Cryptococcus gattii / Q000235:genetics* / Q000302:isolation & purification; D003455:Cryptococcus neoformans / Q000235:genetics* / Q000302:isolation & purification; D004271:DNA, Fungal / Q000032:analysis*; D021903:DNA, Ribosomal Spacer; D006801:Humans; D060888:Real-Time Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
chemical_list: D004271:DNA, Fungal; D021903:DNA, Ribosomal Spacer
keywords: Cryptococcosis; Cryptococcus neoformans/Cryptococcus gattii complex; Molecular diagnosis; Nested PCR