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Multiplex real-time PCR targeting the RNase P RNA gene for detection and identification of Candida species in blood
Linköping University, Faculty of Health Sciences. Linköping University, Department of Molecular and Clinical Medicine, Clinical Microbiology. Östergötlands Läns Landsting, Centre for Laboratory Medicine, Department of Clinical Microbiology.
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2007 (English)In: Journal of Clinical Microbiology, ISSN 0095-1137, Vol. 45, no 3, 874-880 p.Article in journal (Refereed) Published
Abstract [en]

We have developed a single-tube multiplex real-time PCR method for the detection of the eight most common Candida species causing septicemia: Candida albicans, C. dubliniensis, C. famata, C. glabrata, C. guilliermondii, C. krusei, C. parapsilosis, and C. tropicalis. The method developed targets the RNase P RNA gene RPR1. Sequences of this geiie were determined for seven of the Candida species and showed surprisiRgly large sequence variation. C. glabrata was found to have a gene that was five times longer gene than those of the other species, and the nucleotide sequence similarity between C. krusei and C. albicans was as low as 55%. The multiplex PCR contained three probes that enabled the specific detection of C. albicans, C. glabrata, and C. krusei and a fourth probe that allowed the general detection of the remaining species. The method was able to detect 1 to 10 genome copies when the detection limit was tested repeatedly for the four species C. albicans, C. glabrata, C. krusei, and C. guilliermondii. No significant difference in the detection limit was seen when the multiplex format was compared with single-species PCR, i.e., two primers and one probe. The method detected eight clinically relevant Candida species and did not react with other tested non-Candida species or human DNA. The assay was applied to 20 blood samples from nine patients and showed a sensitivity similar to that of culture. Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Place, publisher, year, edition, pages
2007. Vol. 45, no 3, 874-880 p.
National Category
Medical and Health Sciences
URN: urn:nbn:se:liu:diva-38149DOI: 10.1128/JCM.01556-06Local ID: 42045OAI: diva2:258998
Available from: 2009-10-10 Created: 2009-10-10 Last updated: 2011-01-11

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Johansson, Anders
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