Abstract
For Escherichia coli to produce diarrhea in animals it must possess the ability to attach to the epithelial cells of the intestine and to produce enterotoxins. Tests developed to differentiate pathogenic from nonpathogenic E. coli have relied on detection of adherence structures called pili or detection of the toxins. We utilized a monoclonal antibody to K99 pili in an enzyme-linked immunosorbent assay to detect the presence of K99 pili in E. coli isolated from calves. Twenty-three E. coli isolates that were known to be stable toxin positive were all shown to produce K99 pili. A 100% correlation also was shown between the presence of K99 antigen and production of stable toxin by E. coli isolates. Of the 251 isolates, 245 were negative by K99 enzyme-linked immunosorbent assay and stable toxin assay. The other six were positive on both tests. The enzyme-linked immunosorbent assay also was shown to be specific for K99 pili by antibody-blocking assays. The number of E. coli necessary for detecting K99 pili by enzyme-linked immunosorbent assay was determined to be 3.5 X 10(5) bacteria per ml.
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- Arbuckle J. B. The location of Escherichia coli in the pig intestine. J Med Microbiol. 1970 May;3(2):333–340. doi: 10.1099/00222615-3-2-333. [DOI] [PubMed] [Google Scholar]
- Bertschinger H. U., Moon H. W., Whipp S. C. Association of Escherichia coli with the small intestinal epithelium. I. Comparison of enteropathogenic and nonenteropathogenic porcine strains in pigs. Infect Immun. 1972 Apr;5(4):595–605. doi: 10.1128/iai.5.4.595-605.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dean A. G., Ching Y. C., Williams R. G., Harden L. B. Test for Escherichia coli enterotoxin using infant mice: application in a study of diarrhea in children in Honolulu. J Infect Dis. 1972 Apr;125(4):407–411. doi: 10.1093/infdis/125.4.407. [DOI] [PubMed] [Google Scholar]
- Francis D. H., Remmers G. A., DeZeeuw P. S. Production of K88, K99, and 987P antigens by Escherichia coli cultured on synthetic and complex media. J Clin Microbiol. 1982 Jan;15(1):181–183. doi: 10.1128/jcm.15.1.181-183.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Guinée P. A., Jansen W. H. Detection of enterotoxigenicity and attachment factors in Escherichia coli strains of human, porcine and bovine origin; a comparative study. Zentralbl Bakteriol Orig A. 1979 Apr;243(2-3):245–257. [PubMed] [Google Scholar]
- Gyles C. L., Barnum D. A. A heat-labile enterotoxin from strains of Eschericha coli enteropathogenic for pigs. J Infect Dis. 1969 Oct;120(4):419–426. doi: 10.1093/infdis/120.4.419. [DOI] [PubMed] [Google Scholar]
- Hohmann A., Wilson M. R. Adherence of enteropathogenic Escherichia coli to intestinal epithelium in vivo. Infect Immun. 1975 Oct;12(4):866–880. doi: 10.1128/iai.12.4.866-880.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Isaacson R. E., Moon H. W., Schneider R. A. Distribution and virulence of Escherichia coli in the small intestines of calves with and without diarrhea. Am J Vet Res. 1978 Nov;39(11):1750–1755. [PubMed] [Google Scholar]
- Jones G. W., Rutter J. M. Role of the K88 antigen in the pathogenesis of neonatal diarrhea caused by Escherichia coli in piglets. Infect Immun. 1972 Dec;6(6):918–927. doi: 10.1128/iai.6.6.918-927.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kohler E. M. Pathogenesis of neonatal enteric colibacillosis of pigs. J Am Vet Med Assoc. 1972 Feb 15;160(4):574–582. [PubMed] [Google Scholar]
- Merson M. H., Orskov F., Orskov I., Sack R. B., Huq I., Koster F. T. Relationship between enterotoxin production and serotype in enterotoxigenic Escherichia coli. Infect Immun. 1979 Feb;23(2):325–329. doi: 10.1128/iai.23.2.325-329.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Merson M. H., Yolken R. H., Sack R. B., Froehlich J. L., Greenberg H. B., Huq I., Black R. W. Detection of Escherichia coli enterotoxins in stools. Infect Immun. 1980 Jul;29(1):108–113. doi: 10.1128/iai.29.1.108-113.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mills K. W., Phillips R. M., Kelly B. L., Baughman G. L. Using enzyme-linked immunosorbent assay to detect Escherichia coli K88 pili antigens from clinical isolates. Am J Vet Res. 1982 Feb;43(2):365–367. [PubMed] [Google Scholar]
- Mills K. W., Tietze K. L., Phillips R. M. Use of enzyme-linked immunosorbent assay for detection of K88 pili in fecal specimens from swine. Am J Vet Res. 1983 Nov;44(11):2188–2189. [PubMed] [Google Scholar]
- Mooi F. R., de Graaf F. K., van Embden J. D. Cloning, mapping and expression of the genetic determinant that encodes for the K88ab antigen. Nucleic Acids Res. 1979 Mar;6(3):849–865. doi: 10.1093/nar/6.3.849. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Moon H. W., Kohler E. M., Schneider R. A., Whipp S. C. Prevalence of pilus antigens, enterotoxin types, and enteropathogenicity among K88-negative enterotoxigenic Escherichia coli from neonatal pigs. Infect Immun. 1980 Jan;27(1):222–230. doi: 10.1128/iai.27.1.222-230.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Moon H. W., Whipp S. C., Skartvedt S. M. Etiologic diagnosis of diarrheal disease of calves: frequency and methods for detecting enterotoxin and K99 antigen production by Escherichia cola. Am J Vet Res. 1976 Sep;37(9):1025–1029. [PubMed] [Google Scholar]
- Nagy B., Moon H. W., Isaacson R. E. Colonization of porcine small intestine by Escherichia coli: ileal colonization and adhesion by pig enteropathogens that lack K88 antigen and by some acapsular mutants. Infect Immun. 1976 Apr;13(4):1214–1220. doi: 10.1128/iai.13.4.1214-1220.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nakane P. K., Kawaoi A. Peroxidase-labeled antibody. A new method of conjugation. J Histochem Cytochem. 1974 Dec;22(12):1084–1091. doi: 10.1177/22.12.1084. [DOI] [PubMed] [Google Scholar]
- O'Beirne A. J., Cooper H. R. Heterogeneous enzyme immunoassay. J Histochem Cytochem. 1979 Aug;27(8):1148–1162. doi: 10.1177/27.8.383822. [DOI] [PubMed] [Google Scholar]
- Sack D. A., Sack R. B. Test for enterotoxigenic Escherichia coli using Y-1 adrenal cells in miniculture. Infect Immun. 1975 Feb;11(2):334–336. doi: 10.1128/iai.11.2.334-336.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Scherer R. K. Colonial morphology of Escherichia coli on Tergitol-7 medium. Appl Microbiol. 1966 Mar;14(2):152–155. doi: 10.1128/am.14.2.152-155.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Smith H. W., Linggood M. A. Observations on the pathogenic properties of the K88, Hly and Ent plasmids of Escherichia coli with particular reference to porcine diarrhoea. J Med Microbiol. 1971 Nov;4(4):467–485. doi: 10.1099/00222615-4-4-467. [DOI] [PubMed] [Google Scholar]
- Thoen C. O., Mills K., Hopkins M. P. Enzyme-linked protein A: an enzyme-linked immunosorbent assay reagent for detecting antibodies in tuberculous exotic animals. Am J Vet Res. 1980 May;41(5):833–835. [PubMed] [Google Scholar]
- Yolken R. H., Greenberg H. B., Merson M. H., Sack R. B., Kapikian A. Z. Enzyme-linked immunosorbent assay for detection of Escherichia coli heat-labile enterotoxin. J Clin Microbiol. 1977 Nov;6(5):439–444. doi: 10.1128/jcm.6.5.439-444.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- de Graaf F. K., Klemm P., Gaastra W. Purification, characterization, and partial covalent structure of Escherichia coli adhesive antigen K99. Infect Immun. 1981 Sep;33(3):877–883. doi: 10.1128/iai.33.3.877-883.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]