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Applied and Environmental Microbiology, August 2005, p. 4919-4924, Vol. 71, No. 8
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.8.4919-4924.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

SHORT REPORT

Development of PCR Assays Targeting Genes in O-Antigen Gene Clusters for Detection and Identification of Escherichia coli O45 and O55 Serogroups

Chitrita DebRoy,1* Pina M. Fratamico,2 Elisabeth Roberts,1 Michael A. Davis,1 and Yanhong Liu2

Gastroenteric Disease Center, Department of Veterinary Science, The Pennsylvania State University, University Park, Pennsylvania 16802,1 United States Department of Agriculture, Agricultural Research Service, Eastern Regional Research Center, Wyndmoor, Pennsylvania 190382

Received 11 October 2004/ Accepted 21 February 2005

The Escherichia coli O45 O-antigen gene cluster of strain O45:H2 96-3285 was sequenced, and conventional (singleplex), multiplex, and real-time PCR assays were designed to amplify regions in the wzx (O-antigen flippase) and wzy (O-antigen polymerase) genes. In addition, PCR assays targeting the E. coli O55 wzx and wzy genes were designed based on previously published sequences. PCR assays targeting E. coli O45 showed 100% specificity for this serogroup, whereas by PCR assays specific for E. coli O55, 97/102 strains serotyped as E. coli O55 were positive for wzx and 98/102 for wzy. Multiplex PCR assays targeting the E. coli O45 and the E. coli O55 wzx and wzy genes were used to detect the organisms in fecal samples spiked at levels of 106 and 108 CFU/0.2 g feces. Thus, the PCR assays can be used to detect and identify E. coli serogroups O45 and O55.

* Corresponding author. Mailing address: Gastroenteric Disease Center, The Pennsylvania State University, Wiley Lab, Wiley Lane, University Park, PA 16802. Phone: (814) 863-2630. Fax: (814) 863-2167. E-mail: rcd3{at}psu.edu.

Applied and Environmental Microbiology, August 2005, p. 4919-4924, Vol. 71, No. 8
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.8.4919-4924.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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