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Rapid identification of pathogenic yeast isolates by real-time PCR and two-dimensional melting-point analysis

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Abstract

There is a need in the clinical microbiological laboratory for rapid and reliable methods for the universal identification of fungal pathogens. Two different regions of the rDNA gene complex, the highly polymorphic ITS1 and ITS2, were amplified using primers targeting conserved regions of the 18S, 5.8S and 28S genes. After melting-point analysis of the amplified products, the Tm of the two PCR-products were plotted into a spot diagram where all the 14 tested, clinically relevant yeasts separated with good resolution. Real-time amplification of two separate genes, melting-point analysis and two-dimensional plotting of Tm data can be used as a broad-range method for the identification of clinical isolates of pathogenic yeast such as Candida and Cryptococcus spp.

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Acknowledgements

Thanks to N. Kondori, University of Gothenburg, for sending some of the clinical yeast strains used in this study. Thanks also to E. Chryssanthou, Karolinska University Hospital, Stockholm, for the Candida dubliniensis and Candida lusitaniae strains. Thanks to E. Jakobson at the Swedish Institute for Infectious Disease Control Solna, for the work with all the isolates. Also, thanks to all colleagues at the microbiology laboratory at Capio Diagnostik AB in Skövde, for all help and support.

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Authors and Affiliations

  1. Department of Clinical Microbiology, Capio Diagnostik AB, Kärnsjukhuset, Skövde, Sweden

    A. Bergman, B. E. B. Claesson & H. Enroth

  2. Department of Parasitology, Mycology and Environmental Microbiology, Swedish Institute for Infectious Disease Control, Solna, Sweden

    V. Fernandez

  3. School of Life Sciences, University of Skövde, Skövde, Sweden

    K. O. Holmström

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  1. A. Bergman
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  2. V. Fernandez
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  3. K. O. Holmström
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  4. B. E. B. Claesson
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  5. H. Enroth
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Corresponding author

Correspondence to A. Bergman.

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Bergman, A., Fernandez, V., Holmström, K.O. et al. Rapid identification of pathogenic yeast isolates by real-time PCR and two-dimensional melting-point analysis. Eur J Clin Microbiol Infect Dis 26, 813–818 (2007). https://doi.org/10.1007/s10096-007-0369-2

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