論文 - 詳細
| 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 |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | Patent(IFI CLAIMS) |
| 各媒体での言及数の合計 | 2 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| 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 |