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Improved detection methods by genetic and immunological techniques for botulinum C/D and D/C mosaic neurotoxins
- Source :
-
Veterinary Microbiology . Mar2013, Vol. 162 Issue 2-4, p881-890. 10p. - Publication Year :
- 2013
-
Abstract
- Abstract: Clostridium botulinum type C and D strains produce serotype-specific or mosaic botulinum neurotoxin (BoNT). Botulinum C/D and D/C mosaic neurotoxins (BoNT/CD and /DC) are related to avian and bovine botulism, respectively. The two mosaic BoNTs cannot be differentiated from authentic type C and D BoNTs by the conventional serotyping method. In this study, we attempted to establish novel methods for the specific detection of BoNT/CD or/DC. Comparison with nontoxic component genes in type C and D strains revealed that the nucleotide sequence of the ha70 gene is well conserved among either serotype-specific or mosaic BoNT-producing strains. A multiplex PCR method with primers for the light chain of boNT, ntnh, and ha70 gene detection was developed for typing of the boNT gene in type C and D strains. Upon applying this method, twenty-seven type C and D strains, including authentic strains and the isolates from avian and bovine botulism, were successfully divided into type C, C/D mosaic, type D, and D/C mosaic BoNT-producing strains. We then prepared an immunochromatography kit with specific monoclonal antibody showing high binding affinity to each mosaic BoNT. BoNT/CD and /DC in the culture supernatant were detected with limits of detection of 2.5 and 10 LD50, respectively. Furthermore, we confirmed the applicability of the kit for BoNT/DC using crude culture supernatant from a specimen from a bovine suspected of having botulism. These results indicate that the genetic and immunological detection methods are useful for the diagnosis of avian and bovine botulism. [Copyright &y& Elsevier]
Details
- Language :
- English
- ISSN :
- 03781135
- Volume :
- 162
- Issue :
- 2-4
- Database :
- Academic Search Index
- Journal :
- Veterinary Microbiology
- Publication Type :
- Academic Journal
- Accession number :
- 85008626
- Full Text :
- https://doi.org/10.1016/j.vetmic.2012.11.009