Premium
Molecular identification of H istoplasma capsulatum using rolling circle amplification
Author(s) -
Furuie Jason L.,
Sun Jiufeng,
Nascimento Mariana M. F.,
Gomes Renata R.,
WaculiczAndrade Caroline E.,
Sessegolo Gisele C.,
Rodrigues Anderson M.,
GalvãoDias Maria A.,
Camargo Zoilo P.,
QueirozTelles Flávio,
Najafzadeh Mohammad J.,
Hoog Sybren G.,
Vicente Vania A.
Publication year - 2016
Publication title -
mycoses
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.13
H-Index - 69
eISSN - 1439-0507
pISSN - 0933-7407
DOI - 10.1111/myc.12426
Subject(s) - identification (biology) , rolling circle replication , polymerase chain reaction , microbiology and biotechnology , biology , chemistry , genetics , gene , dna replication , botany
Summary 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 (Hc PL 2) allowed specific amplification of H. capsulatum DNA 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.