| Sensitive quantification of Clostridium difficile cells by reverse transcription-quantitative PCR targeting rRNA molecules. | |
| | |
MedLine Citation:
|
PMID: 22582062 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
|
We established a sensitive and accurate quantification system for Clostridium difficile in human intestines, based on rRNA-targeted reverse transcription-quantitative PCR (RT-qPCR). We newly developed a species-specific primer set for C. difficile targeting 23S rRNA gene sequences. Both the vegetative cells and the spores of C. difficile in human feces were quantified by RT-qPCR, with a lower detection limit of 10(2.4) cells/g of feces. In an analysis of the feces of residents (n = 83; age, 85 ± 8 years) and staff (n = 19; age, 36 ± 10 years) at a care facility for the elderly, C. difficile was detected by RT-qPCR in 43% of the residents (average count, log(10) 4.0 ± 2.0 cells/g of feces) and 16% of the staff (average count, log(10) 2.2 ± 0.1 cells/g of feces); these rates were far higher than those detected by qPCR (residents, 19%; staff, 0%) or selective cultivation (residents, 18%; staff, 5%). Another analysis of healthy adults (n = 63; age, 41 ± 11 years) also revealed the significant carriage rate of C. difficile in the intestines (detection rate, 13%; average count, log(10) 4.9 ± 1.2 cells/g of feces). From these results, it was suggested that rRNA-targeted RT-qPCR should be an effective tool for analyzing population levels of C. difficile in the human intestine. |
| | |
Authors:
|
Kazunori Matsuda; Hirokazu Tsuji; Takashi Asahara; Takuya Takahashi; Hiroyuki Kubota; Satoru Nagata; Yuichiro Yamashiro; Koji Nomoto |
Publication Detail:
|
Type: Comparative Study; Evaluation Studies; Journal Article Date: 2012-05-11 |
Journal Detail:
|
Title: Applied and environmental microbiology Volume: 78 ISSN: 1098-5336 ISO Abbreviation: Appl. Environ. Microbiol. Publication Date: 2012 Aug |
Date Detail:
|
Created Date: 2012-07-19 Completed Date: 2012-12-10 Revised Date: 2013-05-20 |
Medline Journal Info:
|
Nlm Unique ID: 7605801 Medline TA: Appl Environ Microbiol Country: United States |
Other Details:
|
Languages: eng Pagination: 5111-8 Citation Subset: IM |
Affiliation:
|
Yakult Central Institute for Microbiological Research, Tokyo, Japan. kazunori-matsuda@yakult.co.jp |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
|
Adult Aged, 80 and over Clostridium difficile / genetics, growth & development* Colony Count, Microbial / methods* DNA Primers / genetics Enterocolitis, Pseudomembranous / epidemiology*, microbiology* Feces / microbiology Humans Intestines / microbiology* Japan / epidemiology RNA, Ribosomal, 23S / genetics* Reverse Transcriptase Polymerase Chain Reaction / methods* |
| Chemical | |
Reg. No./Substance:
|
0/DNA Primers; 0/RNA, Ribosomal, 23S |
| Comments/Corrections | |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: Development of a double-crossover markerless gene deletion system in Bifidobacterium longum: functio...
Next Document: Galacturonic acid inhibits the growth of Saccharomyces cerevisiae on galactose, xylose, and arabinos...