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Article
Characterizing Avian Escherichia coli Isolates with Multiplex Polymerase Chain Reaction
Avian Diseases
  • Jerod A. Skyberg, North Dakota State University
  • Shelley M. Horne, North Dakota State University
  • Catherine W. Giddings, North Dakota State University
  • Richard E. Wooley, University of Georgia
  • Penelope S. Gibbs, North Dakota State University
  • Lisa K. Nolan, Iowa State University
Document Type
Article
Publication Date
10-1-2003
DOI
10.1637/7030
Abstract
Colibacillosis caused by Escherichia coli infections account for significant morbidity and mortality in the poultry industry. Yet, despite the importance of colibacillosis, much about the virulence mechanisms employed by avian E. coli remains unknown. In recent years several genes have been linked to avian E. coli virulence, many of which reside on a large transmissible plasmid. In the present study, a multiplex polymerase chain reaction (PCR) protocol to detect the presence of four of these genes is described. Such a protocol may supplement current diagnostic schemes and provide a rapid means of characterizing the E. coli causing disease in poultry. The targets of this procedure included iss, the increased serum survival gene; tsh, the temperature sensitive hemagglutinin gene; cvi, the ColV immunity gene; and iucC, a gene of the aerobactin operon. Organisms, known for their possession or lack of these genes, were used as a source of the template DNA to develop the multiplex PCR protocol. Identity of the amplicons was confirmed by size, DNA:DNA hybridization with specific gene probes, and DNA sequencing. When the multiplex PCR protocol was used to characterize 10 E. coli isolates incriminated in avian colibacillosis and 10 from the feces of apparently healthy birds, nine of the isolates from apparently healthy birds contained no more than one gene, while the 10th contained all four. Also, eight of the isolates incriminated in colibacillosis contained three or more genes, while the remaining two contained two of the target genes. Interestingly, the isolates of sick birds containing only two of the targeted genes killed the least number of embryos, and the isolate of healthy birds that contained all the genes killed the most embryos among this group. These genes were not found among the non–E. coli isolates tested, demonstrating the procedure's specificity for E. coli. Overall, these results suggest that this protocol might be useful in characterization and study of avian E. coli.
Comments

This article is from Avian Diseases 47, no. 4 (2003): 1441–1447, doi:10.1637/7030.

Copyright Owner
American Association of Avian Pathologists
Language
en
Date Available
May 8, 2013
File Format
application/pdf
Citation Information
Jerod A. Skyberg, Shelley M. Horne, Catherine W. Giddings, Richard E. Wooley, et al.. "Characterizing Avian Escherichia coli Isolates with Multiplex Polymerase Chain Reaction" Avian Diseases Vol. 47 Iss. 4 (2003) p. 1441 - 1447
Available at: http://works.bepress.com/lisa_nolan/57/