Direct and specific identification of Cryptococcus neoformans in biological samples using fluorescently labelled DNA probes.
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.
Martins
M L
ML
Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa, 1349-008 Lisboa, Portugal. luz@ihmt.unl.pt
Ferreira
A S
AS
Sampaio
A
A
Vieira
R
R
Inácio
J
J
D003453:Cryptococcosis / Q000097:blood / Q000134:cerebrospinal fluid / Q000382:microbiology*; D003455:Cryptococcus neoformans / Q000235:genetics / Q000302:isolation & purification*; D015342:DNA Probes / Q000737:chemistry* / Q000235:genetics; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D025202:Molecular Diagnostic Techniques / Q000379:methods
D015342:DNA Probes; D005456:Fluorescent Dyes