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PCR-RFLP as a useful tool for diagnosis of invasivemycoses 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 f...
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Published in: | Electronic Journal of Biotechnology 2015-07, Vol.18 (3) |
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creator | 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 |
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. |
doi_str_mv | 10.1016/j.ejbt.2015.03.012 |
format | article |
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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.</description><identifier>ISSN: 0717-3458</identifier><identifier>EISSN: 0717-3458</identifier><identifier>DOI: 10.1016/j.ejbt.2015.03.012</identifier><language>eng</language><publisher>Universidad Católica de Valparaíso</publisher><subject>Pathogenic fungus ; Polymorphism ; Simultaneous analysis</subject><ispartof>Electronic Journal of Biotechnology, 2015-07, Vol.18 (3)</ispartof><rights>Copyright 2015 - Electronic Journal of Biotechnology</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids></links><search><creatorcontrib>de Sousa, Diego Rayan Teixeira</creatorcontrib><creatorcontrib>Santos, Carla Silvana da Silva</creatorcontrib><creatorcontrib>Wanke, Bodo</creatorcontrib><creatorcontrib>Júnior, Roberto Moreira da Silva</creatorcontrib><creatorcontrib>dos Santos, Marcelo Cordeiro</creatorcontrib><creatorcontrib>Cruz, Kátia Santana</creatorcontrib><creatorcontrib>Monte, Rossicléia Lins</creatorcontrib><creatorcontrib>Nocker, Andreas</creatorcontrib><creatorcontrib>de Souza, João Vicente Braga</creatorcontrib><title>PCR-RFLP as a useful tool for diagnosis of invasivemycoses in a healthcare facility in the North of Brazil</title><title>Electronic Journal of Biotechnology</title><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.</description><subject>Pathogenic fungus</subject><subject>Polymorphism</subject><subject>Simultaneous analysis</subject><issn>0717-3458</issn><issn>0717-3458</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><recordid>eNpNTNtKAzEUDKJgrf6AT_mBXU823WT3UYs3KFqKPpez6YmbJW0kSQv1621RQRiYGzOMXQsoBQh1M5Q0dLmsQNQlyBJEdcJGoIUu5KRuTv_pc3aR0gBQwURPRmyYTxfF4mE255g48m0iu_U8h-C5DZGvHH5sQnKJB8vdZofJ7Wi9NyFROvjDoif0uTcYiVs0zru8Pxa5J_4SYu6Pw7uIX85fsjOLPtHVL4_Z-8P92_SpmL0-Pk9vZ0UnoM0FKmVlS9BZ0ahaGovQIOlWNFq1gmRVQdtJAqtFZ2ttVmoFwthWKbJSWpBjVv78di54t6HlZ3RrjPuliQ6XfyENB4gaZCO_ASkyX98</recordid><startdate>20150710</startdate><enddate>20150710</enddate><creator>de Sousa, Diego Rayan Teixeira</creator><creator>Santos, Carla Silvana da Silva</creator><creator>Wanke, Bodo</creator><creator>Júnior, Roberto Moreira da Silva</creator><creator>dos Santos, Marcelo Cordeiro</creator><creator>Cruz, Kátia Santana</creator><creator>Monte, Rossicléia Lins</creator><creator>Nocker, Andreas</creator><creator>de Souza, João Vicente Braga</creator><general>Universidad Católica de Valparaíso</general><scope>RBI</scope></search><sort><creationdate>20150710</creationdate><title>PCR-RFLP as a useful tool for diagnosis of invasivemycoses in a healthcare facility in the North of Brazil</title><author>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</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-b109t-a66f39e0bf18653cfa08ae79187691e32209b3e0f71bf57cd6d01cf966ef33f03</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Pathogenic fungus</topic><topic>Polymorphism</topic><topic>Simultaneous analysis</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>de Sousa, Diego Rayan Teixeira</creatorcontrib><creatorcontrib>Santos, Carla Silvana da Silva</creatorcontrib><creatorcontrib>Wanke, Bodo</creatorcontrib><creatorcontrib>Júnior, Roberto Moreira da Silva</creatorcontrib><creatorcontrib>dos Santos, Marcelo Cordeiro</creatorcontrib><creatorcontrib>Cruz, Kátia Santana</creatorcontrib><creatorcontrib>Monte, Rossicléia Lins</creatorcontrib><creatorcontrib>Nocker, Andreas</creatorcontrib><creatorcontrib>de Souza, João Vicente Braga</creatorcontrib><collection>Bioline International</collection><jtitle>Electronic Journal of Biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>de Sousa, Diego Rayan Teixeira</au><au>Santos, Carla Silvana da Silva</au><au>Wanke, Bodo</au><au>Júnior, Roberto Moreira da Silva</au><au>dos Santos, Marcelo Cordeiro</au><au>Cruz, Kátia Santana</au><au>Monte, Rossicléia Lins</au><au>Nocker, Andreas</au><au>de Souza, João Vicente Braga</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>PCR-RFLP as a useful tool for diagnosis of invasivemycoses in a healthcare facility in the North of Brazil</atitle><jtitle>Electronic Journal of Biotechnology</jtitle><date>2015-07-10</date><risdate>2015</risdate><volume>18</volume><issue>3</issue><issn>0717-3458</issn><eissn>0717-3458</eissn><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.</abstract><pub>Universidad Católica de Valparaíso</pub><doi>10.1016/j.ejbt.2015.03.012</doi></addata></record> |
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source | SciELO Chile; ScienceDirect Journals |
subjects | Pathogenic fungus Polymorphism Simultaneous analysis |
title | PCR-RFLP as a useful tool for diagnosis of invasivemycoses in a healthcare facility in the North of Brazil |
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