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

Background: The incidence of invasive mycoses is increasing worldwide. PCR-RFLP was applied to the 20identification of 10 reference strains and 90 cultures of agents of invasive mycoses. In addition, the new 21approach was applied to detect fungal agents in 120 biological samples (blood, cerebrospinal fluid and bone 22marrow). PCR-RFLP results were compared with the ones obtained with conventional methods (culture, 23microscopy, and biochemical testing). 24Results: The assays carried out with the reference strains (Candida albicans, Candida parapsilosis, Candida 25tropicalis, Candida krusei, Candida guilliermondii, Cryptococcus neoformans, Cryptococcus gattii and Histoplasma 26capsulatum), demonstrated that the RFLP profiles were correctly predicted by the in silico investigation and 27allowed unequivocal identification of all chosen reference strains. The investigated PCR-RFLP also identified 90 28cultures of agents of invasive mycoses correctly, 2.5 times faster than the conventional assays and was also 29found to be 100% accurate. The RFLP profiles allowed the identification of the etiological agents: C. neoformans 30(n = 3) and C. gattii (n = 1) in CSF samples, H. capsulatum (n = 1) in bone marrow and C. albicans (n = 2) in 31blood cultures. The detection and identification by PCR-RFLP were found to be between two to ten times faster 32than the conventional assays. 33Conclusion:TheresultsshowedthatPCR-RFLPisavaluabletoolfortheidentificationofinvasivemycosesthatcan 34be implemented in hospital laboratories, allowing for a high number of clinical analyses per day. 353637 ©2015Pontificia Universidad Catolica de Valparaiso. Production and hosting by Elsevier B.V. All rights reserved.39404142