Skip to main content
NCBI home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1997 Jan;35(1):228–232. doi: 10.1128/jcm.35.1.228-232.1997

Molecular typing and epidemiological survey of prevalence of Clostridium perfringens types by multiplex PCR.

H S Yoo 1, S U Lee 1, K Y Park 1, Y H Park 1
PMCID: PMC229544  PMID: 8968913

Abstract

Clostridium perfringens has been classified into five toxigenic types (A through E) on the basis of its capability to produce major lethal toxins (alpha, beta, epsilon, and iota toxins). Seroneutralization with mice or guinea pigs has been used to type each toxin, but this conventional method has some disadvantages. Therefore, we used a molecular biological technique to type the bacterium in the present study. A multiplex PCR was developed for this purpose. This method has several advantages in comparison with seroneutralization with mice or guinea pigs. By this method, we also investigated the most prevalent type(s) of the organism in Korean calves, piglets, and chickens showing clinical symptoms such as diarrhea, enterotoxemia, and necrotic enteritis. Only type A was isolated from calves and chickens, while type C (2 of 14 isolates), in addition to type A, was isolated from piglets. These results suggested that seroneutralization could be replaced by our new method and that type A of C. perfringens is the most prevalent type in livestock in Korea.

Full Text

The Full Text of this article is available as a PDF (271.0 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Al-Sheikhly F., Truscott R. B. The interaction of Clostridium perfringens and its toxins in the production of necrotic enteritis of chickens. Avian Dis. 1977 Apr-Jun;21(2):256–263. [PubMed] [Google Scholar]
  2. Awad M. M., Bryant A. E., Stevens D. L., Rood J. I. Virulence studies on chromosomal alpha-toxin and theta-toxin mutants constructed by allelic exchange provide genetic evidence for the essential role of alpha-toxin in Clostridium perfringens-mediated gas gangrene. Mol Microbiol. 1995 Jan;15(2):191–202. doi: 10.1111/j.1365-2958.1995.tb02234.x. [DOI] [PubMed] [Google Scholar]
  3. Bhown A. S., Habeerb A. F. Structural studies on epsilon-prototoxin of Clostridium perfringens type D. Localization of the site of tryptic scission necessary for activation to epsilon-toxin. Biochem Biophys Res Commun. 1977 Oct 10;78(3):889–896. doi: 10.1016/0006-291x(77)90506-x. [DOI] [PubMed] [Google Scholar]
  4. CRAIG J. P., MILES A. A. Some properties of the iota-toxin of Clostridium welchii, including its action on capillary permeability. J Pathol Bacteriol. 1961 Apr;81:481–493. doi: 10.1002/path.1700810221. [DOI] [PubMed] [Google Scholar]
  5. Collins J. E., Bergeland M. E., Bouley D., Ducommun A. L., Francis D. H., Yeske P. Diarrhea associated with Clostridium perfringens type A enterotoxin in neonatal pigs. J Vet Diagn Invest. 1989 Oct;1(4):351–353. doi: 10.1177/104063878900100414. [DOI] [PubMed] [Google Scholar]
  6. Daube G., China B., Simon P., Hvala K., Mainil J. Typing of Clostridium perfringens by in vitro amplification of toxin genes. J Appl Bacteriol. 1994 Dec;77(6):650–655. doi: 10.1111/j.1365-2672.1994.tb02815.x. [DOI] [PubMed] [Google Scholar]
  7. Hatheway C. L. Toxigenic clostridia. Clin Microbiol Rev. 1990 Jan;3(1):66–98. doi: 10.1128/cmr.3.1.66. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hunter S. E., Brown J. E., Oyston P. C., Sakurai J., Titball R. W. Molecular genetic analysis of beta-toxin of Clostridium perfringens reveals sequence homology with alpha-toxin, gamma-toxin, and leukocidin of Staphylococcus aureus. Infect Immun. 1993 Sep;61(9):3958–3965. doi: 10.1128/iai.61.9.3958-3965.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hunter S. E., Clarke I. N., Kelly D. C., Titball R. W. Cloning and nucleotide sequencing of the Clostridium perfringens epsilon-toxin gene and its expression in Escherichia coli. Infect Immun. 1992 Jan;60(1):102–110. doi: 10.1128/iai.60.1.102-110.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Itodo A. E., Adesiyun A. A., Adekeye J. O., Umoh J. U. Toxin-types of Clostridium perfringens strains isolated from sheep, cattle and paddock soils in Nigeria. Vet Microbiol. 1986 Jun;12(1):93–96. doi: 10.1016/0378-1135(86)90045-3. [DOI] [PubMed] [Google Scholar]
  11. Johnson S., Echeverria P., Taylor D. N., Paul S. R., Coninx R., Sakurai J., Eampokalap B., Jimakorn P., Cooke R. A., Lawrence G. W. Enteritis necroticans among Khmer children at an evacuation site in Thailand. Lancet. 1987 Aug 29;2(8557):496–500. doi: 10.1016/s0140-6736(87)91803-4. [DOI] [PubMed] [Google Scholar]
  12. Jolivet-Reynaud C., Moreau H., Alouf J. E. Purification of alpha toxin from Clostridium perfringens: phospholipase C. Methods Enzymol. 1988;165:91–94. doi: 10.1016/s0076-6879(88)65017-8. [DOI] [PubMed] [Google Scholar]
  13. Katayama S., Matsushita O., Minami J., Mizobuchi S., Okabe A. Comparison of the alpha-toxin genes of Clostridium perfringens type A and C strains: evidence for extragenic regulation of transcription. Infect Immun. 1993 Feb;61(2):457–463. doi: 10.1128/iai.61.2.457-463.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lawrence G., Brown R., Bates J., Saul A., Davis M., Spark R., Anian G. An affinity technique for the isolation of Clostridium perfringens type C from man and pigs in Papua New Guinea. J Appl Bacteriol. 1984 Oct;57(2):333–338. doi: 10.1111/j.1365-2672.1984.tb01398.x. [DOI] [PubMed] [Google Scholar]
  15. Lawrence G., Cooke R. Experimental pigbel: the production and pathology of necrotizing enteritis due to Clostridium welchii type C in the guinea-pig. Br J Exp Pathol. 1980 Jun;61(3):261–271. [PMC free article] [PubMed] [Google Scholar]
  16. Lawrence G., Walker P. D. Pathogenesis of enteritis necroticans in Papula New Guinea. Lancet. 1976 Jan 17;1(7951):125–126. doi: 10.1016/s0140-6736(76)93160-3. [DOI] [PubMed] [Google Scholar]
  17. Murray M. G., Thompson W. F. Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res. 1980 Oct 10;8(19):4321–4325. doi: 10.1093/nar/8.19.4321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Murrell T. G., Egerton J. R., Rampling A., Samels J., Walker P. D. The ecology and epidemiology of the pig-bel syndrome in man in New Guinea. J Hyg (Lond) 1966 Sep;64(3):375–396. doi: 10.1017/s0022172400040663. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Nagahama M., Kobayashi K., Ochi S., Sakurai J. Enzyme-linked immunosorbent assay for rapid detection of toxins from Clostridium perfringens. FEMS Microbiol Lett. 1991 Nov 1;68(1):41–44. doi: 10.1016/0378-1097(91)90392-n. [DOI] [PubMed] [Google Scholar]
  20. Naylor R. D., Martin P. K., Sharpe R. T. Detection of Clostridium perfringens epsilon toxin by ELISA. Res Vet Sci. 1987 Mar;42(2):255–256. [PubMed] [Google Scholar]
  21. Niilo L. Clostridium perfringens Type C Enterotoxemia. Can Vet J. 1988 Aug;29(8):658–664. [PMC free article] [PubMed] [Google Scholar]
  22. Niilo L. Enterotoxigenic Clostridium perfringens type A isolated from intestinal contents of cattle, sheep and chickens. Can J Comp Med. 1978 Jul;42(3):357–363. [PMC free article] [PubMed] [Google Scholar]
  23. Ohsaka A., Tsuchiya M., Oshio C., Miyairi M., Suzuki K., Yamakawa Y. Aggregation of platelets in the mesenteric microcirculation of the rat induced by alpha-toxin (phospholipase C) of Clostridium perfringens. Toxicon. 1978;16(4):333–341. doi: 10.1016/0041-0101(78)90153-8. [DOI] [PubMed] [Google Scholar]
  24. Perelle S., Gibert M., Boquet P., Popoff M. R. Characterization of Clostridium perfringens iota-toxin genes and expression in Escherichia coli. Infect Immun. 1993 Dec;61(12):5147–5156. doi: 10.1128/iai.61.12.5147-5156.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Popoff M. R., Jestin A. Enteropathogenicity of purified Clostridium perfringens enterotoxin in the pig. Am J Vet Res. 1985 Oct;46(10):2147–2148. [PubMed] [Google Scholar]
  26. Smith L. D. Virulence factors of Clostridium perfringens. Rev Infect Dis. 1979 Mar-Apr;1(2):254–262. doi: 10.1093/clinids/1.2.254. [DOI] [PubMed] [Google Scholar]
  27. Songer J. G. Clostridial enteric diseases of domestic animals. Clin Microbiol Rev. 1996 Apr;9(2):216–234. doi: 10.1128/cmr.9.2.216. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Stiles B. G., Wilkins T. D. Purification and characterization of Clostridium perfringens iota toxin: dependence on two nonlinked proteins for biological activity. Infect Immun. 1986 Dec;54(3):683–688. doi: 10.1128/iai.54.3.683-688.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Stubbings D. P. Clostridium perfringens enterotoxaemia in two young horses. Vet Rec. 1990 Oct 27;127(17):431–431. [PubMed] [Google Scholar]
  30. Sugahara T., Takahashi T., Yamaya S., Ohsaka A. Vascular permeability increase by alpha-toxin (phospholipase C) of Clostridium perfringens. Toxicon. 1977;15(1):81–87. doi: 10.1016/0041-0101(77)90074-5. [DOI] [PubMed] [Google Scholar]
  31. Truscott R. B., Al-Sheikhly F. Reproduction and treatment of necrotic enteritis in broilers. Am J Vet Res. 1977 Jun;38(6):857–861. [PubMed] [Google Scholar]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES