Quantitative real time PCR assays for the enumeration of Saccharomyces cerevisiae and Saccharomyces sensu stricto complex in human feces

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Title
Quantitative real time PCR assays for the enumeration of Saccharomyces cerevisiae and Saccharomyces sensu stricto complex in human feces
Author(s)
Ho Won Chang; Young Do Nam; Y Sung; Kyoung-Ho Kim; SeongWoon Roh; Jung-Hoon Yoon; K G An; Jin-Woo Bae
Bibliographic Citation
Journal of Microbiological Methods, vol. 71, no. 3, pp. 191-201
Publication Year
2007
Abstract
There have been an increasing number of reports of yeast systemic infection involving Saccharomyces cerevisiae strains. The development of a rapid and reliable diagnostic tool is therefore warranted in order to explore the distribution of S. cerevisiae as an opportunistic pathogen in humans. In this study, we designed and validated five primer sets targeting the 26S rRNA gene of S. cerevisiae and the S. sensu stricto complex using 26 yeast strains. Among them, two sets of primers specifically amplified the 26S rRNA gene and the ITS region of S. cerevisiae strains, and three sets were specific for amplifying the same genes in the S. sensu stricto complex. After determining the optimal conditions of two primer pairs for quantitative real time PCR, human fecal samples were analyzed to examine the distribution of S. cerevisiae and the S. sensu stricto complex. It was possible to detect a single cell of S. cerevisiae in environmental sample. Qualitative PCR revealed that out of eleven fecal samples tested, one sample contained S. cerevisiae and four samples contained the S. sensu stricto complex. Quantitative real time PCR revealed that the target gene copy numbers of S. cerevisiae and the S. sensu stricto complex were 0.84 and 2.44 respectively, in 1 ng of DNA from the bulk fecal community.
Keyword
human fecesreal time PCRsaccharomyces cerevisiaesaccharomyces sensu stricto
ISSN
0167-7012
Publisher
Elsevier
DOI
http://dx.doi.org/10.1016/j.mimet.2007.08.013
Type
Article
Appears in Collections:
1. Journal Articles > Journal Articles
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