Abstract
Although the etiology of porcine proliferative enteritis is not understood, the consistent presence of intracellular Campylobacter-like organisms (CLOs) in proliferating pig intestinal epithelial cells suggests that the organism is involved in the disease process. In order to obtain information about this organism, we generated and characterized specific DNA probes to the intracellular CLO which was purified without culturing. Intracellular CLOs were isolated from mucosa by homogenization, filtration, and absorption to wheat germ agglutinin-Sepharose. The DNA was purified, and a CLO genomic library was constructed. The specificity of recombinant plasmids was confirmed by both dot blot hybridization and Southern analysis of normal and diseased mucosa, as well as of a variety of Campylobacter species. Several of the CLO-specific probes hybridized with porcine mucosa obtained from pigs with proliferative enteritis but not with nondiseased mucosa. The probes hybridized equally with mucosa or DNA obtained from each of the two clinical forms of proliferative enteritis, i.e., proliferative hemorrhagic enteropathy and porcine intestinal adenomatosis. The CLO-specific probes failed to hybridize with any of the commonly isolated porcine Campylobacter species, including Campylobacter hyointestinalis, C. mucosalis, and C. coli. Therefore, the intracellular CLO of porcine proliferative enteritis may be an as yet unidentified or uncultured species.
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- Boosinger T. R., Thacker H. L., Armstrong C. H. Campylobacter sputorum subsp mucosalis and Campylobacter hyointestinalis infections in the intestine of gnotobiotic pigs. Am J Vet Res. 1985 Oct;46(10):2152–2156. [PubMed] [Google Scholar]
- Chang K., Kurtz H. J., Ward G. E., Gebhart C. J. Immunofluorescent demonstration of Campylobacter hyointestinalis and Campylobacter sputorum subsp mucosalis in swine intestines with lesions of proliferative enteritis. Am J Vet Res. 1984 Apr;45(4):703–710. [PubMed] [Google Scholar]
- Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
- Gebhart C. J., Murtaugh M. P., Lin G. F., Ward G. E. Species-specific DNA probes for Campylobacter species isolated from pigs with proliferative enteritis. Vet Microbiol. 1990 Sep;24(3-4):367–379. doi: 10.1016/0378-1135(90)90184-w. [DOI] [PubMed] [Google Scholar]
- Gebhart C. J., Ward G. E., Chang K., Kurtz H. J. Campylobacter hyointestinalis (new species) isolated from swine with lesions of proliferative ileitis. Am J Vet Res. 1983 Mar;44(3):361–367. [PubMed] [Google Scholar]
- Gebhart C. J., Ward G. E., Murtaugh M. P. Species-specific cloned DNA probes for the identification of Campylobacter hyointestinalis. J Clin Microbiol. 1989 Dec;27(12):2717–2723. doi: 10.1128/jcm.27.12.2717-2723.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Holmes D. S., Quigley M. A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem. 1981 Jun;114(1):193–197. doi: 10.1016/0003-2697(81)90473-5. [DOI] [PubMed] [Google Scholar]
- Lawson G. H., Rowland A. C., MacIntyre N. Demonstration of a new intracellular antigen in porcine intestinal adenomatosis and hamster proliferative ileitis. Vet Microbiol. 1985 Jun;10(4):303–313. doi: 10.1016/0378-1135(85)90001-x. [DOI] [PubMed] [Google Scholar]
- McOrist S., Boid R., Lawson G. H. Antigenic analysis of Campylobacter species and an intracellular Campylobacter-like organism associated with porcine proliferative enteropathies. Infect Immun. 1989 Mar;57(3):957–962. doi: 10.1128/iai.57.3.957-962.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McOrist S., Boid R., Lawson G. H., McConnell I. Monoclonal antibodies to intracellular campylobacter-like organisms of the porcine proliferative enteropathies. Vet Rec. 1987 Oct 31;121(18):421–422. doi: 10.1136/vr.121.18.421. [DOI] [PubMed] [Google Scholar]
- Ohya T., Kubo M., Watase H. Campylobacter species isolated from swine with lesions of proliferative enteritis. Nihon Juigaku Zasshi. 1985 Apr;47(2):285–294. doi: 10.1292/jvms1939.47.285. [DOI] [PubMed] [Google Scholar]
- Owen R. J., Borman P. A rapid biochemical method for purifying high molecular weight bacterial chromosomal DNA for restriction enzyme analysis. Nucleic Acids Res. 1987 Apr 24;15(8):3631–3631. doi: 10.1093/nar/15.8.3631. [DOI] [PMC free article] [PubMed] [Google Scholar]