I´m looking for suitable RT-PCR assay to detect and distinguish Candida albicans, C. glabrata and C. krusei. Preferably assay compatible with Biorad CFX96 cycler and already used in clinical settings. We already tried assay from White et al, JMM 2003 and Khlif et al, CMI 2009 but it didn't work as good as we expect. I will be grateful for any advice or suggestions.
Best regard
Jan
Dear Jan,
In recent years the taxonomy of many pathogenic Candida species has been revised due to the description of closely related cryptic species difficult to identify. For example C. albicans is now recognized as cryptic species complex comprising C. dubliniensis and C. africana. Also C. glabrata is a cryptic complex and C. nivariensis and C. bracarensis are two phylogenetically related species. I don't know RT-PCR_based assays for their identification but you can use my 2 simple published PCR protocols for their proper identification/discrimination:
1) Romeo et al. A multiplex PCR protocol for rapid identification of Candida glabrata and its phylogenetically related species Candida nivariensis and Candida bracarensis. J Microbiol Methods. 2009;79:117-20.
2) Romeo and Criseo. First molecular method for discriminating between Candida africana, Candida albicans, and Candida dubliniensis by using hwp1 gene. Diagn Microbiol Infect Dis. 2008;62:230-3. (this is the sole PCR-based method that allow the identification of C. africana).
I hope that this can help you.
If you have any question regarding molecular identification of Candida species, please do not hesitate to contact me ([email protected])
Best regards
Orazio
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