PCR-RFLP as a useful tool for diagnosis of invasive mycoses in a healthcare facility in the North of Brazil

• Diego Rayan Teixeira de Sousa ^[1] ; Carla Silvana da Silva Santos ^[2] ; Bodo Wanke ^[3] ; Roberto Moreira da Silva Júnior ^[1] ; Marcelo Cordeiro dos Santos ^[2] ; Kátia Santana Cruz ^[2] ; Rossicléia Lins Monte ^[2] ; Andreas Nocker ^[2] ; João Vicente Braga de Souza ^[1]
  1. [1] Universidade do Estado do Amazonas

     Universidade do Estado do Amazonas

     Brasil

     
  2. [2] Fundação de Medicina Tropical Dr. Heitor Vieira Dourado Departamento de Pesquisa Laboratório de Micologia
  3. [3] Fundação Oswaldo Cruz Departamento de Pesquisa Laboratório de Micologia

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• Localización: Electronic Journal of Biotechnology, ISSN-e 0717-3458, Vol. 18, Nº. 3, 2015, págs. 231-235
• Idioma: inglés
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  • 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.