Molecular identification of Histoplasma capsulatum using rolling circle amplification

Molecular identification of Histoplasma capsulatum using rolling circle amplification

Author Furuie, Jason L. Google Scholar
Sun, Jiufeng Google Scholar
do Nascimento, Mariana M. F. Google Scholar
Gomes, Renata R. Google Scholar
Waculicz-Andrade, Caroline E. Google Scholar
Sessegolo, Gisele C. Google Scholar
Rodrigues, Anderson M. Autor UNIFESP Google Scholar
Galvao-Dias, Maria A. Google Scholar
de Camargo, Zoilo P. Autor UNIFESP Google Scholar
Queiroz-Telles, Flavio Google Scholar
Najafzadeh, Mohammad J. Google Scholar
de Hoog, Sybren G. Google Scholar
Vicente, Vania A. Google Scholar
Abstract Histoplasmosis is a systemic fungal disease that occurs worldwide, causing symptomatic infection mostly in immunocompromised hosts. Etiological agent is the dimorphic fungus, Histoplasma capsulatum, which occurs in soil contaminated with bird or bat droppings. Major limitation in recognition of H. capsulatum infections is the low awareness, since other diseases may have similar symptomatology. The molecular methods have gained importance because of unambiguous diagnostic ability and efficiency. The aim of this study was to develop and evaluate a padlock probe in view of rolling circle amplification (RCA) detection method which targets ITS (Internal Transcribed Spacer) rDNA of H. capsulatum enabling rapid and specific detection of the fungus in clinical samples. Two padlock probes were designed and one of these (HcPL2) allowed specific amplification of H. capsulatumDNA while no cross-reactivity was observed with fungi used as negative controls. This method proved to be effective for H. capsulatum specific identification and demonstrated to be faster than the traditional method of microbiological identification.
Keywords Histoplasmosis
Histoplasma capsulatum
diagnosis
padlock probe
RCARapid Identification
Genetic Diversity
Species Complex
Pcr
Sequence
Regions
Polymorphism
Brazil
Assay
Dna
Language English
Sponsor Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Date 2016
Published in Mycoses. Hoboken, v. 59, n. 1, p. 12-19, 2016.
ISSN 0933-7407 (Sherpa/Romeo, impact factor)
Publisher Wiley-Blackwell
Extent 12-19
Origin http://dx.doi.org/10.1111/myc.12426
Access rights Closed access
Type Article
Web of Science ID WOS:000367095200003
URI http://repositorio.unifesp.br/handle/11600/49692

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