Rapid and simple detection of Clostridium botulinum types A and B by loop-mediated isothermal amplification
Article first published online: 2 FEB 2009
© 2009 The Authors. Journal compilation © 2009 The Society for Applied Microbiology
Journal of Applied Microbiology
Volume 106, Issue 4, pages 1252–1259, April 2009
How to Cite
Sakuma, T., Kurosaki, Y., Fujinami, Y., Takizawa, T. and Yasuda, J. (2009), Rapid and simple detection of Clostridium botulinum types A and B by loop-mediated isothermal amplification. Journal of Applied Microbiology, 106: 1252–1259. doi: 10.1111/j.1365-2672.2008.04084.x
- Issue published online: 9 MAR 2009
- Article first published online: 2 FEB 2009
- 2008/0439: received 13 March 2008, revised 19 September 2008 and accepted 26 September 2008
- Clostridium botulinum;
- detection method;
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.