Abstract
Attaching and effacing Escherichia coli is a new causal agent of diarrhea in calves. Its major virulence factors are the intimin protein, encoded by the eaeA gene, and the Shiga-like toxins, encoded by slt genes. Because the sequences of these genes are available, we selected specific primers to amplify each virulence gene so as to develop a new identification test based on multiplex amplification of virulence-associated genes. Of 30 tested strains, 14 were eaeA+, 15 were eaeA+ slt-I+, 1 was eaeA+ slt-I+ slt-II+, and 1 was eaeA+ slt-II+. The method proved in our hands to be fast and specific and in perfect correlation with the hybridization method.
Full Text
The Full Text of this article is available as a PDF (633.1 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Beebakhee G., Louie M., De Azavedo J., Brunton J. Cloning and nucleotide sequence of the eae gene homologue from enterohemorrhagic Escherichia coli serotype O157:H7. FEMS Microbiol Lett. 1992 Feb 1;70(1):63–68. doi: 10.1016/0378-1097(92)90563-4. [DOI] [PubMed] [Google Scholar]
- Boyer H. W., Roulland-Dussoix D. A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol. 1969 May 14;41(3):459–472. doi: 10.1016/0022-2836(69)90288-5. [DOI] [PubMed] [Google Scholar]
- Brian M. J., Frosolono M., Murray B. E., Miranda A., Lopez E. L., Gomez H. F., Cleary T. G. Polymerase chain reaction for diagnosis of enterohemorrhagic Escherichia coli infection and hemolytic-uremic syndrome. J Clin Microbiol. 1992 Jul;30(7):1801–1806. doi: 10.1128/jcm.30.7.1801-1806.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chanter N., Morgan J. H., Bridger J. C., Hall G. A., Reynolds D. J. Dysentery in gnotobiotic calves caused by atypical Escherichia coli. Vet Rec. 1984 Jan 21;114(3):71–71. doi: 10.1136/vr.114.3.71. [DOI] [PubMed] [Google Scholar]
- Fratamico P. M., Sackitey S. K., Wiedmann M., Deng M. Y. Detection of Escherichia coli O157:H7 by multiplex PCR. J Clin Microbiol. 1995 Aug;33(8):2188–2191. doi: 10.1128/jcm.33.8.2188-2191.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gannon V. P., Rashed M., King R. K., Thomas E. J. Detection and characterization of the eae gene of Shiga-like toxin-producing Escherichia coli using polymerase chain reaction. J Clin Microbiol. 1993 May;31(5):1268–1274. doi: 10.1128/jcm.31.5.1268-1274.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jackson M. P., Newland J. W., Holmes R. K., O'Brien A. D. Nucleotide sequence analysis of the structural genes for Shiga-like toxin I encoded by bacteriophage 933J from Escherichia coli. Microb Pathog. 1987 Feb;2(2):147–153. doi: 10.1016/0882-4010(87)90106-9. [DOI] [PubMed] [Google Scholar]
- Jerse A. E., Yu J., Tall B. D., Kaper J. B. A genetic locus of enteropathogenic Escherichia coli necessary for the production of attaching and effacing lesions on tissue culture cells. Proc Natl Acad Sci U S A. 1990 Oct;87(20):7839–7843. doi: 10.1073/pnas.87.20.7839. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Knutton S., Baldwin T., Williams P. H., McNeish A. S. Actin accumulation at sites of bacterial adhesion to tissue culture cells: basis of a new diagnostic test for enteropathogenic and enterohemorrhagic Escherichia coli. Infect Immun. 1989 Apr;57(4):1290–1298. doi: 10.1128/iai.57.4.1290-1298.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Konowalchuk J., Speirs J. I., Stavric S. Vero response to a cytotoxin of Escherichia coli. Infect Immun. 1977 Dec;18(3):775–779. doi: 10.1128/iai.18.3.775-779.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Levine M. M. Escherichia coli that cause diarrhea: enterotoxigenic, enteropathogenic, enteroinvasive, enterohemorrhagic, and enteroadherent. J Infect Dis. 1987 Mar;155(3):377–389. doi: 10.1093/infdis/155.3.377. [DOI] [PubMed] [Google Scholar]
- Lin Z., Kurazono H., Yamasaki S., Takeda Y. Detection of various variant verotoxin genes in Escherichia coli by polymerase chain reaction. Microbiol Immunol. 1993;37(7):543–548. doi: 10.1111/j.1348-0421.1993.tb01675.x. [DOI] [PubMed] [Google Scholar]
- Mainil J. G., Bex F., Jacquemin E., Pohl P., Couturier M., Kaeckenbeeck A. Prevalence of four enterotoxin (STaP, STaH, STb, and LT) and four adhesin subunit (K99, K88, 987P, and F41) genes among Escherichia coli isolates from cattle. Am J Vet Res. 1990 Feb;51(2):187–190. [PubMed] [Google Scholar]
- Mainil J. G., Duchesnes C. J., Whipp S. C., Marques L. R., O'Brien A. D., Casey T. A., Moon H. W. Shiga-like toxin production and attaching effacing activity of Escherichia coli associated with calf diarrhea. Am J Vet Res. 1987 May;48(5):743–748. [PubMed] [Google Scholar]
- Mainil J. G., Jacquemin E. R., Kaeckenbeeck A. E., Pohl P. H. Association between the effacing (eae) gene and the Shiga-like toxin-encoding genes in Escherichia coli isolates from cattle. Am J Vet Res. 1993 Jul;54(7):1064–1068. [PubMed] [Google Scholar]
- Newland J. W., Neill R. J. DNA probes for Shiga-like toxins I and II and for toxin-converting bacteriophages. J Clin Microbiol. 1988 Jul;26(7):1292–1297. doi: 10.1128/jcm.26.7.1292-1297.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- O'Brien A. D., Holmes R. K. Shiga and Shiga-like toxins. Microbiol Rev. 1987 Jun;51(2):206–220. doi: 10.1128/mr.51.2.206-220.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- O'Brien A. D., LaVeck G. D. Purification and characterization of a Shigella dysenteriae 1-like toxin produced by Escherichia coli. Infect Immun. 1983 May;40(2):675–683. doi: 10.1128/iai.40.2.675-683.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Scotland S. M., Smith H. R., Willshaw G. A., Rowe B. Vero cytotoxin production in strain of Escherichia coli is determined by genes carried on bacteriophage. Lancet. 1983 Jul 23;2(8343):216–216. doi: 10.1016/s0140-6736(83)90192-7. [DOI] [PubMed] [Google Scholar]
- Yu J., Kaper J. B. Cloning and characterization of the eae gene of enterohaemorrhagic Escherichia coli O157:H7. Mol Microbiol. 1992 Feb;6(3):411–417. doi: 10.1111/j.1365-2958.1992.tb01484.x. [DOI] [PubMed] [Google Scholar]