Molecular identification of H istoplasma capsulatum using rolling circle amplification

• Published in: Mycoses Vol. 59; no. 1; pp. 12 - 19
• Main Authors: Furuie, Jason L., Sun, Jiufeng, do Nascimento, Mariana M. F., Gomes, Renata R., Waculicz‐Andrade, Caroline E., Sessegolo, Gisele C., Rodrigues, Anderson M., Galvão‐Dias, Maria A., de Camargo, Zoilo P., Queiroz‐Telles, Flávio, Najafzadeh, Mohammad J., de Hoog, Sybren G., Vicente, Vania A.
• Format: Journal Article
• Language: English
• Published: 01-01-2016
• ISSN: 0933-7407; 1439-0507
• DOI: 10.1111/myc.12426

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.