論文 - 詳細
RRC ID | 17413 |
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著者 | Sakuma T, Kurosaki Y, Fujinami Y, Takizawa T, Yasuda J. |
タイトル | Rapid and simple detection of Clostridium botulinum types A and B by loop-mediated isothermal amplification. |
ジャーナル | J Appl Microbiol |
Abstract |
AIMS:To develop a convenient and rapid detection method for toxigenic Clostridium botulinum types A and B using a loop-mediated isothermal amplification (LAMP) method. METHODS AND RESULTS:The LAMP primer sets for the type A or B botulinum neurotoxin gene, BoNT/A or BoNT/B, were designed. To determine the specificity of the LAMP assay, a total of 14 C. botulinum strains and 17 other Clostridium strains were tested. The assays for the BoNT/A or BoNT/B gene detected only type A or B C. botulinum strains, respectively, but not other types of C. botulinum or strains of other Clostridium species. Using purified chromosomal DNA, the sensitivity of LAMP for the BoNT/A or BoNT/B gene was 1 pg or 10 pg of DNA per assay, respectively. The assay times needed to detect 1 ng of DNA were only 23 and 22 min for types A and B, respectively. In food samples, the detection limit per reaction was one cell for type A and 10 cells for type B. CONCLUSIONS:The LAMP is a sensitive, specific and rapid detection method for C. botulinum types A and B. SIGNIFICANCE AND IMPACT OF THE STUDY:The LAMP assay would be useful for detection of C. botulinum in environmental samples. |
巻・号 | 106(4) |
ページ | 1252-9 |
公開日 | 2009-4-1 |
DOI | 10.1111/j.1365-2672.2008.04084.x |
PII | JAM4084 |
PMID | 19187148 |
MeSH | Base Sequence Clostridium botulinum / genetics* Clostridium botulinum / isolation & purification DNA Primers / genetics DNA, Bacterial / analysis* DNA, Bacterial / genetics Food Microbiology Molecular Sequence Data Nucleic Acid Amplification Techniques / methods Plasmids / genetics Reverse Transcriptase Polymerase Chain Reaction / methods Sensitivity and Specificity |
IF | 3.066 |
引用数 | 11 |
WOS 分野 | BIOTECHNOLOGY & APPLIED MICROBIOLOGY MICROBIOLOGY |
リソース情報 | |
一般微生物 | JCM 1290 JCM 1296 JCM 1382 JCM 1385 JCM 1391 JCM 1406 JCM 1409 JCM 1410 JCM 1416 JCM 1417 JCM 3816 JCM 7835 JCM 7850 JCM 11011 |