Comparison of molecular techiniques for identification of fusarium species from clinical samples

Author(s)

Marcela de Souza

• Document type: Master's Dissertation
• Press: Campinas, SP.
• Institution: Universidade Estadual de Campinas (UNICAMP). Faculdade de Ciências Médicas
• Defense date: 2014-02-13

Examining board members

Plinio Trabasso; Mariângela Ribeiro Resende; Luci Correa

• Advisor: Plinio Trabasso

Abstract

This study aimed to compare the performance of the techniques of DNA microarray, loop mediated isothermal amplification (LAMP), real-time PCR and DNA sequencing for identification of species of Fusarium as agents of bloodstream infection, isolated from patients with hematologic malignancies cared for at the university hospital of State University of Campinas and the university hospital of University of Sao Paulo, in a retrospective cross study. In the study, 21 Fusarium isolates from blood were selected, being 17 samples of mycology collection of the Laboratory for Research on Fungi (LIF), Faculty of Medical Sciences, Campinas (FCM-Unicamp) and three samples of mycology collection of the Laboratory of Microbiology of FMUSP. The techniques of DNA microarray, LAMP and Real-Time PCR identified 17 of the 21 samples as Fusarium solani, and 4 isolates as Fusarium non-solani complex, showing a high performance, a 100% agreement among them and a sensitivity of 100% for the DNA microarray, LAMP and real-time PCR methods and a 100% specificity for the LAMP method. These data were validated by the DNA sequencing technique, which identified 17 isolates as Fusarium solani complex and not four as Fusarium non-solani complex, being three Fusarium napiforme and one Fusarium oxysporum, agreeing with the findings in the three methodologies applied in this work. The LAMP technique proved to be more accessible than the techniques of DNA microarray and real-time PCR to be faster, indicating a promising methodology to be used in routine diagnosis of filamentous fungi, aiding in proper therapy rapid and specific manner, thereby providing an improvement to the care of patients (AU)

• FAPESP's process: 11/16205-5 - Identification of Fusarium species from clinical specimens from Campinas State University Hospital through the techniques of DNA microarray and loop-mediated isothermal amplification (LAMP).
• Grantee: Marcela de Souza
• Support Opportunities: Scholarships in Brazil - Master