RRC ID 17413
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