Molecules and Cells

  • 제24권3호
  • /
  • Pages.329-337
  • /
  • 2007
  • /
  • 1016-8478(pISSN)
  • /
  • 0219-1032(eISSN)
한국분자세포생물학회 (Korean Society for Molecular and Cellular Biology)
권호 표지

Development of Enrichment Semi-nested PCR for Clostridium botulinum types A, B, E, and F and Its Application to Korean Environmental Samples

  • Shin, Na-Ri (Division of High-Risk Pathogen Research, Center for Infectious Diseases, Korean National Institute of Health) ;
  • Yoon, So-Yeon (Division of High-Risk Pathogen Research, Center for Infectious Diseases, Korean National Institute of Health) ;
  • Shin, Ji-Hun (Division of High-Risk Pathogen Research, Center for Infectious Diseases, Korean National Institute of Health) ;
  • Kim, Yun Jeong (Division of High-Risk Pathogen Research, Center for Infectious Diseases, Korean National Institute of Health) ;
  • Rhie, Gi-eun (Division of High-Risk Pathogen Research, Center for Infectious Diseases, Korean National Institute of Health) ;
  • Kim, Bong Su (Division of High-Risk Pathogen Research, Center for Infectious Diseases, Korean National Institute of Health) ;
  • Seong, Won Keun (Division of Biosafety Evaluation and Control, Center for Infectious Diseases, Korean National Institute of Health) ;
  • Oh, Hee-Bok (Division of High-Risk Pathogen Research, Center for Infectious Diseases, Korean National Institute of Health)
  • 투고 : 2007.04.13
  • 심사 : 2007.07.19
  • 발행 : 2007.12.31
⟨ 이전 논문 다음 논문 ⟩

초록

An enrichment semi-nested PCR procedure was developed for detection of Clostridium botulinum types A, B, E, and F. It was applied to sediment samples to examine the prevalence of C. botulinum in the Korean environment. The first pair of primers for the semi-nested PCR was designed using a region shared by the types A, B, E, and F neurotoxin gene sequences, and the second round employed four nested primers complementary to the BoNT/A, /B, /E, and /F encoding genes for simultaneous detection of the four serotypes. Positive results were obtained from the PCR analysis of five of 44 sediments (11%) collected from Yeong-am Lake in Korea; all were identified as deriving from type B neurotoxin (bontb) genes. Two of the C. botulinum type B organisms were isolated, and their bontb genes sequenced. The deduced amino acid sequences of BoNT/B showed 99.5 and 99.8% identity with the amino acid sequence of accession no. AB084152. Our data suggest that semi-nested PCR is a useful tool for detecting C. botulinum in sediments, and renders it practicable to conduct environmental surveys.

키워드

과제정보

연구 과제 주관 기관 : National Institute of Health

참고문헌

  1. Aranda, E., Rodriguez, M. M., Asensio, M. A., and Cordoba, J. J. (1997) Detection of Clostridium botulinum types A, B, E and F in foods by PCR and DNA probe. Lett. Appl. Microbiol. 24, 186−190 https://doi.org/10.1046/j.1472-765X.1997.00374.x
  2. Austin, J. W. and Blanchfield, B. (1997) Detection of Clostridium botulinum and its toxins in suspect foods and clinical specimens. Polyscience Publications publ. MFHPB-16, Quebec, Canada
  3. Borland, E. D., Moryson, C. J., and Smith, G. R. (1977) Avian botulism and the high prevalence of Clostridium botulinum in the Norflok Broads. Vet. Res. 100, 106−109
  4. Campbell, K. D., Collins, M. D., and East, A. K. (1993) Gene probes for identification of the botulinal neurotoxin gene and specific identification of neurotoxin types B, E, and F. J. Clin. Microbiol. 31, 2255−2262
  5. Carlin, F., Broussolle, V., Perelle, S., Litman, S., and Fach, P. (2004) Prevalence of Clostridium botulinum in food raw materials used in REPFEDs manufactured in France. Int. J. Food Microbiol. 91, 141−145
  6. Center for disease control. (1998) Botulism in the United States, 1899−1996. Handbook for epidemiologists, clinicians, and laboratory workers. DHEW Publ. No. (CDC) 74−8279, Washington, DC, plus additional reports by CDC at annual meetings of the Interagency Botulism Research Coordinating Committee (IBRCC)
  7. Chaffer, M., Baum, M., Grinberg, K., Molad, T., and Elad, D. (2006) Application of PCR for detection Clostridium botulinum type D in bovine samples. J. Vet. Med. 53, 45−47
  8. Collins, M. D. and East, A. K. (1998) Phylogeny and taxonomy of the food-borne pathogen Clostridium botulinum and its neurotoxins. J. Appl. Microbiol. 84, 5−17
  9. Dahlenborg, M., Borch, E., and Radstrom, P. (2001) Development of a combined selection and enrichment PCR procedure for Clostridium botulinum types B, E, and F and its use to determine prevalence in fecal samples from slaughtered pigs. Appl. Environ. Microbiol. 67, 4781−4 https://doi.org/10.1128/AEM.67.2.814-820.2001
  10. Dahlenborg, M., Borch, E., and Radstrom, P. (2003) Prevalence of Clostridium botulinum types B, E, and F in faecal samples from Swedish cattle. Int. J. Food Microbiol. 82, 105−110
  11. Fach, P., Gilbert, M., Griffais, R., Guillou, J. P., and Poppff, M. R. (1993) Polymerase chain reaction for the rapid identification of Clostridium botulinum type A strains and detection in food samples. J. Appl. Bacteriol. 75, 234−239
  12. Fach, P., Gibert, M., Griffais, R., Guillou, J. P., and Popoff, M. R. (1995) PCR and gene probe identification of botulinum neurotoxin A-, B-, E-, F-, and G-producing Clostridium spp. and evaluation in food samples. Appl. Environ. Microbiol. 61, 389−392
  13. Fach, P., Perelle, S., Dilasser, F., Grout, J., Dargaignaratz, C., et al. (2002) Detection by PCR-enzyme-linked immunosorbent assay of Clostridium botulinum in fish and environmental samples from a coastal area in northern France. Appl. Environ. Microbiol. 68, 5870−5876
  14. Franciosa, G., Ferreira, J. L., and Hatheway, C. L. (1994) Detection of type A, B, and E botulism neurotoxin genes in Clostridium botulinum and other Clostridium species by PCR: evidence of unexpressed type B toxin genes in type A toxigenic organisms. J. Clin. Microbiol. 32, 1911−1917
  15. Galey, F. D., Terra, R., Walker, R., Adaska, J., Etchebarne. M. A., et al. (2000) Type C botulism in dairy cattle from feed contaminated with a dead cat. J. Vet. Diagn. Invest. 12, 204−209
  16. Geissler, E. and Moon J. E., ed. (1999) Biological and toxin weapons: Research, development and use from the Middle ages to 1945. New York, NY: Oxford University Press; Sipri Chemical & Biological Warfare Studies No. 18
  17. Hatheway, C. L. (1990) Toxigenic clostridia. Clin. Microbiol. Rev. 3, 66−98
  18. Hielm, S., Hyytia, E., Andersin, A. B., and Korkeala, H. (1998) A high prevalence of Clostridium botulinum type E in Finnish freshwater and Baltic sea sediment samples. J. Appl. Microbiol. 84, 133−137
  19. Holdeman, L. V. and Brooks, J. B. (1970) Variation among strains of Clostridium botulinum and related clostridia. In Proceedings of the First U.S.-Japan Conference of Toxic Microorganisms ed. Herzberg, M. pp. 278−286. Washington DC: US Government Printing Office
  20. Hyytia, E., Hielm, S., and Korkeala, H. (1998) Prevalence of Clostridium botulinum type E in Finnish and fishery products. Epidemiol. Infect. 120, 245−250
  21. Kimura, B., Kawasaki, S., Nakano, H., and Fujii, T. (2001) Rapid, quantitative PCR monitoring of growth of Clostridium botulinum type E in modified-atmosphere-packaged fish. Appl. Environ. Microbiol. 67, 206−216
  22. Lalitha, K. V. and Gopakumar, K. (2000) Distribution and ecology of Clostridium botulinum in fish and aquatic environments of a tropical region. Food Microbiol. 17, 535−541
  23. Lindstrom, M. and Korkeala, H. (2006) Laboratory Diagnostics of botulism. Clin. Microbiol. Rev. 19, 298−314
  24. Marion, W., O'Meara, T. E., Riddle, G. D., and Berkhoff, H. A. (1983) Prevalence of Clostridium botulinum type C in substrates of phosphate-mine settling ponds and implications for epizootics of avian botulism. J. Wildlife Dis. 19, 302−307
  25. Martin, S. (2003) Clostridium botulinum type D intoxication in a dairy herd in Ontario. Can. Vet. J. 44, 493−495
  26. Nol, P., Rocke, T. E., Gross, K., and Yuill, T. M. (2004) Prevalence of neurotoxic Clostridium botulinum type C in the gastrointestinal tracts of Tilapia (Oreochromis mossambicus) in the salton sea. J. Wildlife Dis. 40, 414−419
  27. Peck, M. W., Fairbairn, D. A., and Lund, B. M. (1992) The effect of recovery medium on the estimated heat-inactivation of spores of non-proteolytic Clostridium botulinum. Lett. Appl. Microbiol. 15, 146−151
  28. Rocke, T. E., Smith, S. R., and Nashold, S. W. (1998) Preliminary evaluation of a simple in vitro test for the diagnosis of type C botulism in wild birds. J. Wildlife Dis. 34, 744−751
  29. Schocken-Iturrino, R. P., Carneiro, M. C., Kato, E., Sorbara, J. O. B., Rossi, O. D., et al. (1999) Study of the presence of the spores of Clostridium botulinum in honey in Brazil. FEMS. Immunol. Med. Microbiol. 24, 379−382
  30. Shin, N. R., Shin, J. H., Chun, J. H., Yoon, S. Y., Kim, B. S., et al. (2006) Determination of neurotoxin gene expression in Clostridium botulinum type A by quantitative RT-PCR. Mol. Cells 22, 336−342
  31. Zechmeister, T. C., Kirschner, A. K., Fuchsberger, M., Gruber, S. G., Suess, B., et al. (2005) Prevalence of botulinum neurotoxin C1 and its corresponding gene in environmental samples from low and high risk avian botulism areas. ALTEX 22, 185-195