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Title: PCR-RFLP as a useful tool for diagnosis of invasivemycoses in a healthcare facility in the North of Brazil
Authors: Sousa, Diego Rayan Teixeira de
Silva Santos, Carla Silvana da
Wanke, Bodo
Silva, Roberto Moreira da
dos Santos, Marcelo Cordeiro
Cruz, Kátia Santana
Monte, Rossicléia Lins
Nocker, Andreas
Souza, João Vicente Braga de
Keywords: Blood
Cerebrospinal Fluid
Polymerase Chain Reaction
Candida Guilliermondii
Candida Parapsilosis
Conventional Methods
Cryptococcus Neoformans
Detection And Identifications
Healthcare Facility
Pathogenic Fungi
Simultaneous Analysis
Ajellomyces Capsulatus
Biochemical Analysis
Blood Analysis
Bone Marrow Examination
Candida Albicans
Candida Parapsilosis
Candida Tropicalis
Cerebrospinal Fluid Examination
Controlled Study
Cryptococcus Gattii
Cryptococcus Neoformans
Diagnostic Accuracy
Diagnostic Sensibility
Diagnostic Test Accuracy Study
Fungal Cell Culture
Fungal Strain
Fungus Identification
Major Clinical Study
Meyerozyma Guilliermondii
Middle Aged
Pichia Kudriavzevii
Polymerase Chain Reaction
Quality Control Procedures
Polymorphism, Restriction Fragment Length
Sensitivity And Specificity
Systemic Mycosis
Young Adult
Ajellomyces Capsulatus
Candida Albicans
Candida Parapsilosis
Candida Tropicalis
Cryptococcus Gattii
Filobasidiella Neoformans
Issatchenkia Orientalis
Pichia Guilliermondii
Issue Date: 2015
metadata.dc.publisher.journal: Electronic Journal of Biotechnology
metadata.dc.relation.ispartof: Volume 18, Número 3, Pags. 231-235
Abstract: Background: The incidence of invasive mycoses is increasing worldwide. PCR-RFLP was applied to the identification of 10 reference strains and 90 cultures of agents of invasive mycoses. In addition, the new approach was applied to detect fungal agents in 120 biological samples (blood, cerebrospinal fluid and bone marrow). PCR-RFLP results were compared with the ones obtained with conventional methods (culture, microscopy, and biochemical testing). Results: The assays carried out with the reference strains (Candida albicans, Candida parapsilosis, Candida tropicalis, Candida krusei, Candida guilliermondii, Cryptococcus neoformans, Cryptococcus gattii and Histoplasma capsulatum), demonstrated that the RFLP profiles were correctly predicted by the in silico investigation and allowed unequivocal identification of all chosen reference strains. The PCR-RFLP also identified 90 cultures of agents of invasive mycoses correctly, 2.5 times faster than the conventional assays. Evaluating PCR-RFLP with biological samples it was observed that the PCR was found to be 100% accurate and the RFLP profiles allowed the identification of the etiological agents: C. neoformans (n = 3) and C. gattii (n = 1) in CSF samples, H. capsulatum (n = 1) in bone marrow and C. albicans (n = 2) in blood cultures. The detection and identification by PCR-RFLP were found to be between two to ten times faster than the conventional assays. Conclusion: The results showed that PCR-RFLP is a valuable tool for the identification of invasive mycoses that can be implemented in hospital laboratories, allowing for a high number of clinical analyses per day. © 2015 Pontificia Universidad Católica de Valparaíso. Production and hosting by Elsevier B.V. All rights reserved.
metadata.dc.identifier.doi: 10.1016/j.ejbt.2015.03.012
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