| Autor: |
Sakuma, T., Kurosaki, Y., Fujinami, Y., Takizawa, T., Yasuda, J. |
| Předmět: |
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| Zdroj: |
Journal of Applied Microbiology; Apr2009, Vol. 106 Issue 4, p1252-1259, 8p, 1 Diagram, 5 Charts, 1 Graph |
| Abstrakt: |
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. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
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