Genotypic Characterization of Clostridium botulinum Strains Producing Type A Neurotoxin Complexes by Random Amplified Polymorphic DNA and PCR-RFLP Analysis
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- ISHIMURA Katsuyuki
- Department of Biological Science, Hiroshima City Institute of Public Health
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- NAKANO Hiroyuki
- Laboratory of Food Microbiology and Hygiene, Graduate School of Biosphere Science, Hiroshima University
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- KAYASHIMA Takayuki
- Department of Biological Science, Hiroshima City Institute of Public Health
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- OGINO Takeo
- Department of Biological Science, Hiroshima City Institute of Public Health
Bibliographic Information
- Other Title
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- RAPDおよびPCR-RFLP法によるA型毒素産生ボツリヌス菌の遺伝子型鑑
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Abstract
In order to genotypically characterize Clostridium botulinum type A strains, we examined 23 strains from various origins (food-borne botulism, infant botulism, honey, honey-related materials and soil), by random amplified polymorphic DNA (RAPD) analysis and characterized the type A botulinum neurotoxin complex genes by PCR and PCR-RFLP analysis. By RAPD analysis, C. botulinum strains associated with infant botulism in Japan and a strain 804-1H, isolated from Brazilian honey, were clearly differentiated from the other strains examined.PCR and PCR-RFLP analysis were performed for the neurotoxin complex genes (type Aneurotoxin (BoNT/A), type B neurotoxin (BoNT/B), nontoxic-nonhemagglutinin (NTNH), hemaggulutinin (HA), and p47 protein). All of 23 strains tested gave positive amplification for BoNT/A and NTNH genes, and the PCR-RFLP analysis differentiated six strains associated with infant botulism in Japan, a strain from Brazilian honey and a strain AF84 (type 2) from other strains (type 1). For HA gene, the type 1 strains gave amplification while the type 2 strains gave no amplification. For BoNT/B gene, the type 1 strains (a strain Renkon and three American infant botulism strains) gave amplification (silent B strains). For p47 gene, the type 2 strains and the silent B strains were positive. The combinations of the PCR of HA, p47, and BoNT/B genes and the PCR-RFLP of BoNT/A, NTNH genes demonstrated three genotypes (A 1, A2, and A3) of neurotoxin complex genes. Thus, these PCR-based methods are useful for simple and rapid differentiation of the BoNT-producing C. botulinum strains.
Journal
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- Japanese Journal of Food Microbiology
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Japanese Journal of Food Microbiology 22 (4), 148-154, 2005
Japanese Society of Food Microbiology
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Details 詳細情報について
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- CRID
- 1390001204425574016
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- NII Article ID
- 130004103566
- 10016965107
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- NII Book ID
- AN10552871
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- ISSN
- 18825982
- 13408267
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- NDL BIB ID
- 7803851
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- Text Lang
- en
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- Data Source
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- JaLC
- NDL
- Crossref
- CiNii Articles
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- Abstract License Flag
- Disallowed