Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: March 30, 2005
Publication Date: August 15, 2005
Citation: Fratamico, P.M., Debroy, C., Miyamoto, T., Liu, Y. 2005. Detection of Enterohemorrhagic Escherichia Coli 0145 by PCR Targeting Genes in the E. Coli 0145 Antigen Gene Cluster and the Shiga Toxin 1 and Shiga Toxin 2 Genes. Association of Official Analytical Chemists. Abstract. Paper # P-611. Technical Abstract: Enterohemorrhagic Escherichia coli serogroup O145 is an important cause of hemorrhagic colitis and hemolytic uremic syndrome worldwide. Cattle and other animals are potential reservoirs for this pathogen. To develop PCR assays for detection and identification of E. coli O145, the DNA sequence of the O antigen gene cluster of this E. coli serogroup was determined, and 15 open reading frames were identified. The wzx (O antigen flippase) and wzy (O antigen polymerase) genes in the O145 O antigen gene cluster were selected as targets for development of PCR assays specific for this serogroup. In addition, a multiplex PCR assay targeting the E. coli O145 wzx and wzy genes and the Shiga toxin 1 (stx1) and Shiga toxin 2 (stx2) genes was developed and used to detect Shiga toxin-producing E. coli O145 in seeded ground beef initially inoculated with ca. 2 CFU/25g of meat. The PCR assays can be employed to identify E. coli serogroup O145 and to detect low levels of the pathogen in ground beef and potentially in other foods and in environmental samples.