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Detection and identification of Candida spp. from bloodstream infections of critically ill pediatric patients: validation of a multiplex PCR

Grant number: 10/02626-6
Support Opportunities:Scholarships in Brazil - Master
Effective date (Start): August 01, 2010
Effective date (End): July 31, 2012
Field of knowledge:Health Sciences - Medicine - Maternal and Child Health
Principal Investigator:Gilda Maria Barbaro Del Negro
Grantee:Cleison Ledesma Taira
Host Institution: Faculdade de Medicina (FM). Universidade de São Paulo (USP). São Paulo , SP, Brazil


Disseminated candidiasis is an important cause of morbidity and mortality in immunocompromised and critically ill pediatric patients admitted at intensive care units. Increased use of immunosupressive drugs, indwelling catheters, broad-spectrum antibiotics, parenteral nutrition and other predisposing conditions have contributed to the increment of Candida bloodstream infections. Laboratory diagnosis of candidemia is troublesome due to the lack of blood cultures sensitivity. Detection of Candida DNA by polymerase chain reaction (PCR) in tiny blood volumes may provide a more rapid and reliable candidemia detection in critically ill children. The aim of this study is to validate a multiplex PCR technique for detection and identification of seven Candida species in pediatric patients bloodstream at risk to develop fungemia, and comparing results with those obtained by blood cultures. Blood samples collected from 30 newborns admitted at Neonatal Intensive Care Unit and 30 patients admitted at Pediatric Intensive Care Unit of Instituto da Criança-HCFMUSP, presenting with predisposing conditions to develop candidemia will be analyzed. A control group composed of 30 non infected children, admitted at the pediatric surgery unit of the same hospital to undergo small interventions such as hernia and fimosis correction will be included. The limit of detection and the specificity of the multiplex PCR will be determined employing species-specific primers designed in the present study. DNA extracted from blood samples will be submitted to amplifications and PCR products analyzed by agarose gel electrophoresis. If PCR positive results are not confirmed by blood cultures, DNA sequencing will be performed. Agreement between both methods will be assessed by the McNemar test, adopting a level of significance of 5%. (AU)

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
TAIRA, CLEISON LEDESMA; OKAY, THELMA SUELY; DELGADO, ARTUR FIGUEIREDO; JURFEST RIVERO CECCON, MARIA ESTHER; GOTTARDO DE ALMEIDA, MARGARETE TERESA; BARBARO DEL NEGRO, GILDA MARIA. A multiplex nested PCR for the detection and identification of Candida species in blood samples of critically ill paediatric patients. BMC INFECTIOUS DISEASES, v. 14, . (10/09715-4, 10/02626-6)
Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)
TAIRA, Cleison Ledesma. Validation of a PCR multiplex assay for detection and identification of Candida spp. in bloodstream infections of critically ill pediatric patients. 2013. Master's Dissertation - Universidade de São Paulo (USP). Faculdade de Medicina (FM/SBD) São Paulo.

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