RRC ID 34104
著者 Sato N, Seo G, Benno Y.
タイトル Development of strain-specific PCR primers for quantitative detection of Bacillus mesentericus strain TO-A in human feces.
ジャーナル Biol Pharm Bull
Abstract Strain-specific polymerase chain reaction (PCR) primers for detection of Bacillus mesentericus strain TO-A (BM TO-A) were developed. The randomly amplified polymorphic DNA (RAPD) technique was used to produce potential strain-specific markers. A 991-bp RAPD marker found to be strain-specific was sequenced, and two primer pairs specific to BM TO-A were constructed based on this sequence. In addition, we explored a more specific DNA region using inverse PCR, and designed a strain-specific primer set for use in real-time quantitative PCR (qPCR). These primer pairs were tested against 25 Bacillus subtilis strains and were found to be strain-specific. After examination of the detection limit and linearity of detection of BM TO-A in feces, the qPCR method and strain-specific primers were used to quantify BM TO-A in the feces of healthy volunteers who had ingested 3×10(8) colony forming unit (CFU) of BM TO-A per day in tablets. During the administration period, BM TO-A was detected in the feces of all 24 subjects, and the average number of BM TO-A detected using the culture method and qPCR was about 10(4.8) and 10(5.8) cells per gram of feces, respectively. Using the qPCR method, BM TO-A was detected in the feces of half of the subjects 3 d after withdrawal, and was detected in the feces of only one subject 1 week after withdrawal. These results suggest that the qPCR method using BM TO-A strain-specific primers is useful for the quantitative detection of this strain in feces.
巻・号 37(1)
ページ 123-9
公開日 2014-1-1
DOI 10.1248/bpb.b13-00641
PII DN/JST.JSTAGE/bpb/b13-00641
PMID 24389488
MeSH Bacillus / genetics* Colon / microbiology* Colony Count, Microbial DNA Primers* DNA, Bacterial / analysis* Feces / microbiology* Genetic Markers Humans Random Amplified Polymorphic DNA Technique* Real-Time Polymerase Chain Reaction Sequence Analysis, DNA / methods*
IF 1.863
引用数 1
WOS 分野 PHARMACOLOGY & PHARMACY
リソース情報
一般微生物 JCM 9070 JCM 10629 JCM 20014 JCM 20073 JCM 20096 JCM 20105 JCM 20197 JCM 20230 JCM 20333 JCM 20336 JCM 20354 JCM 20520