Document Detail


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:  2013-06-25    
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
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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

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