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

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 fl...

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Autores principales:	Sousa,Diego Rayan Teixeira de, Santos,Carla Silvana da Silva, Wanke,Bodo, Silva Júnior,Roberto Moreira da, Santos,Marcelo Cordeiro dos, Cruz,Kátia Santana, Monte,Rossicléia Lins, Nocker,Andreas, Souza,João Vicente Braga de
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2015
Materias:	Pathogenic fungus Polymorphism Simultaneous analysis
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582015000300014
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spelling	oai:scielo:S0717-345820150003000142015-06-26PCR-RFLP as a useful tool for diagnosis of invasive mycoses in a healthcare facility in the North of BrazilSousa,Diego Rayan Teixeira deSantos,Carla Silvana da SilvaWanke,BodoSilva Júnior,Roberto Moreira daSantos,Marcelo Cordeiro dosCruz,Kátia SantanaMonte,Rossicléia LinsNocker,AndreasSouza,João Vicente Braga de Pathogenic fungus Polymorphism Simultaneous analysis 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.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.18 n.3 20152015-05-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582015000300014en10.1016/j.ejbt.2015.03.012
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	Pathogenic fungus Polymorphism Simultaneous analysis
spellingShingle	Pathogenic fungus Polymorphism Simultaneous analysis Sousa,Diego Rayan Teixeira de Santos,Carla Silvana da Silva Wanke,Bodo Silva Júnior,Roberto Moreira da Santos,Marcelo Cordeiro dos Cruz,Kátia Santana Monte,Rossicléia Lins Nocker,Andreas Souza,João Vicente Braga de PCR-RFLP as a useful tool for diagnosis of invasive mycoses in a healthcare facility in the North of Brazil
description	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.
author	Sousa,Diego Rayan Teixeira de Santos,Carla Silvana da Silva Wanke,Bodo Silva Júnior,Roberto Moreira da Santos,Marcelo Cordeiro dos Cruz,Kátia Santana Monte,Rossicléia Lins Nocker,Andreas Souza,João Vicente Braga de
author_facet	Sousa,Diego Rayan Teixeira de Santos,Carla Silvana da Silva Wanke,Bodo Silva Júnior,Roberto Moreira da Santos,Marcelo Cordeiro dos Cruz,Kátia Santana Monte,Rossicléia Lins Nocker,Andreas Souza,João Vicente Braga de
author_sort	Sousa,Diego Rayan Teixeira de
title	PCR-RFLP as a useful tool for diagnosis of invasive mycoses in a healthcare facility in the North of Brazil
title_short	PCR-RFLP as a useful tool for diagnosis of invasive mycoses in a healthcare facility in the North of Brazil
title_full	PCR-RFLP as a useful tool for diagnosis of invasive mycoses in a healthcare facility in the North of Brazil
title_fullStr	PCR-RFLP as a useful tool for diagnosis of invasive mycoses in a healthcare facility in the North of Brazil
title_full_unstemmed	PCR-RFLP as a useful tool for diagnosis of invasive mycoses in a healthcare facility in the North of Brazil
title_sort	pcr-rflp as a useful tool for diagnosis of invasive mycoses in a healthcare facility in the north of brazil
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2015
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582015000300014
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PCR-RFLP as a useful tool for diagnosis of invasive mycoses in a healthcare facility in the North of Brazil

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