Abstract
Fluorescence in situ hybridisation (FISH) is a suitable technique for the rapid, reliable and cultivation-independent identification of microbial pathogens. This study describes the development of fluorescently labelled rRNA-targeted oligonucleotides and a FISH assay to detect and identify Cryptococcus neoformans in culture and biological samples. All C. neoformans reference and clinical isolates gave positive signals with the specific oligonucleotide probes, whereas all non-target yeast species gave negative reactions with the same probes. The assay was also successfully applied to the detection of C. neoformans cells in cerebrospinal samples from patients with clinical diagnosis of cryptococcosis. The described FISH-based assay revealed to be practical, sensitive and specific for the detection and identification of C. neoformans yeasts.
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Acknowledgements
We thank Maria José Salgado and Lurdes Monteiro (Hospital de Santa Maria, Lisbon), Zélia Saraiva and Teresa Tendeiro (Instituto Português de Oncologia, Lisbon), and Virgínia Lopes (Hospital de Santo António, Oporto) for providing the yeast isolates and/or clinical samples. The authors would also like to acknowledge Prof. Isabel Spencer-Martins, prematurely deceased on June 15th 2008, for her valuable suggestions on the present work.
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Martins, M.L., Ferreira, A.S., Sampaio, A. et al. Direct and specific identification of Cryptococcus neoformans in biological samples using fluorescently labelled DNA probes. Eur J Clin Microbiol Infect Dis 29, 571–576 (2010). https://doi.org/10.1007/s10096-010-0897-z
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DOI: https://doi.org/10.1007/s10096-010-0897-z