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Electronic Journal of Biotechnology
Universidad Católica de Valparaíso
ISSN: 0717-3458
Vol. 18, No. 3, 2015, pp. 231-235
Bioline Code: ej15038
Full paper language: English
Document type: Research Article
Document available free of charge

Electronic Journal of Biotechnology, Vol. 18, No. 3, 2015, pp. 231-235

 en PCR-RFLP as a useful tool for diagnosis of invasivemycoses in a healthcare facility in the North of Brazil
de Sousa, Diego Rayan Teixeira; Santos, Carla Silvana da Silva; Wanke, Bodo; Júnior, Roberto Moreira da Silva; dos Santos, Marcelo Cordeiro; Cruz, Kátia Santana; Monte, Rossicléia Lins; Nocker, Andreas & de Souza, João Vicente Braga

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 check for this species in other resources , Candida parapsilosis check for this species in other resources , Candida tropicalis check for this species in other resources , Candida krusei check for this species in other resources , Candida guilliermondii check for this species in other resources , Cryptococcus neoformans check for this species in other resources , Cryptococcus gattii check for this species in other resources and Histoplasma capsulatum check for this species in other resources ), 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.

Keywords
Pathogenic fungus; Polymorphism; Simultaneous analysis

 
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