U. Evertsson et al., Detection and identification of fungi in blood using broad-range 28S rDNA PCR amplification and species-specific hybridisation, APMIS, 108(5), 2000, pp. 385-392
The aim of the present study was to develop a PCR-based method to detect an
d identify fungi directly from human venous blood. We used broad-range PCR
primers that targeted a part of the large subunit 28S rRNA genes. To obtain
species-specific hybridisation probes, type strains of Candida albicans, C
. glabrata, C. krusei, C. parapsilosis, C. tropicalis and Cryptococcus neof
ormans were PCR amplified, and the amplicons were analysed by gene sequenci
ng. Based on the sequence analysis, species-specific probes that targeted v
ariable regions were designed and used in hybridisation analyses. Between 2
to 10 fungal cells/ml of spiked blood samples could be detected and correc
tly identified to species. We applied the technique to blood samples obtain
ed from two patients with or two patients without verified candidaemia. The
three samples of candidaemia patients were correctly identified to species
level, and those of the negative patients remained negative. This method i
s a potential tool for diagnosis of systemic invasive candidiasis.