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Characterization of paracoccidioides brasiliensis by ft-ir spectroscopy and nanotechnology

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Date
2016
Author
Ferreira, Isabelle
Ferreira-Strixino, Juliana
Castilho, Maiara L.
Campos, Claudia Barbosa Ladeira de [UNIFESP]
Tellez, Claudio
Raniero, Leandro
Type
Artigo
ISSN
1386-1425
Is part of
Spectrochimica Acta Part A-Molecular And Biomolecular Spectroscopy
DOI
10.1016/j.saa.2015.07.061
Metadata
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Abstract
Paracoccidioides brasiliensis, the etiological agent of paracoccidioidomycosis, is a dimorphic fungus existing as mycelia in the environment (or at 25 degrees C in vitro) and as yeast cells in the human host (or at 37 degrees C in vitro). Because mycological examination of lesions in patients frequently is unable to show the presence of the fungus and serological tests can misdiagnose the disease with other mycosis, the development of new approach's for molecular identification of P. brasiliensis spurges is needed. This study describes the use of a gold nanoprobe of a known gene sequence of P. brasiliensis as a molecular tool to identify P. brasiliensis by regular polymerase chain reaction (PCR) associated with a colorimetric methods. This approach is suitable for testing in remote areas because it does not require any further step than gene amplification, being safer and cheaper than electrophoresis methods. The proposed test showed a color change of the PCR reaction mixture from red to blue in negative samples, whereas the solution remains red in positive samples. We also performed a Fourier Transform Infrared (FT-IR) Spectroscopy analysis to characterize and compare the chemical composition between yeast and mycelia forms, which revealed biochemical differences between these two forms. The analysis of the spectra showed that differences were distributed in chemical bonds of proteins, lipids and carbohydrates. The most prominent difference between both forms was vibration modes related to 1,3-beta-glucan usually found in mycelia and 1,3-alpha-glucan found in yeasts and also chitin forms. In this work, we introduce FT-IR as a new method suitable to reveal overall differences that biochemically distinguish each form of P. brasiliensis that could be additionally used to discriminate biochemical differences among a single form under distinct environmental conditions. (C) 2015 Elsevier B.V. All rights reserved.
Citation
Spectrochimica Acta Part A-Molecular And Biomolecular Spectroscopy. Oxford, v. 152, p. 397-403, 2016.
Keywords
Paracoccidioides Brasiliensis
Yeast
Mycelia
Gold Nanoprobes
Ft-IrGold Nanoparticles
Cell-Wall
Glycoprotein Gp43
Gene-Expression
Dna
Chitin
Yeast
Identification
Antigen
Protein
Sponsorship
FAPESP [2009/09559-5]
CNPq [301066/2009-4, 301022/2012-7]
URI
http://repositorio.unifesp.br/handle/11600/49195
Collections
  • ICT - Artigos [439]

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