Direct and specific identification of in biological samples using fluorescently labelled DNA probes

M. L. Martins (1, 2, 3) , A. S. Ferreira (1, 4) , A. Sampaio (4) , R. Vieira (5) , J. Inácio (6)

1 IHMT - Instituto de Higiene e Medicina Tropical
2 Centro de Recursos Microbiológicos (CREM), Departamento de Ciências da Vida, Faculdade de Ciências e Tecnologia
3 Lab. Micologia
4 Centro de Investigação e de Tecnologias Agroambientais e Biológicas (CITAB)
5 Serviço de Dermatologia
6 Laboratório Nacional de Investigação Veterinária (LNIV–INRB, I.P.)

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 in culture and biological samples. All 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 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 yeasts.

Mots clés: Life Sciences

M. L. Martins, A. S. Ferreira, A. Sampaio, R. Vieira, J. Inácio. Direct and specific identification of in biological samples using fluorescently labelled DNA probes. European Journal of Clinical Microbiology and Infectious Diseases, 2010, 29 (5), pp.571-576. ⟨10.1007/s10096-010-0897-z⟩. ⟨hal-00575680⟩