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Journal of Clinical Microbiology, August 2004, p. 3799-3804, Vol. 42, No. 8
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.8.3799-3804.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Identification of Escherichia coli O114 O-Antigen Gene Cluster and Development of an O114 Serogroup-Specific PCR Assay

Lu Feng,1,2,3 Wei Wang,1 Jiang Tao,1 Hongjie Guo,1,2,3 Gladys Krause,4 Lothar Beutin,4 and Lei Wang1,2,3*

TEDA School of Biological Sciences and Biotechnology,1 Tianjin State Laboratory of Microbial Functional Genomics, TEDA College, Nankai University,2 Tianjin Biochip Corporation, TEDA, Tianjin 300457, People's Republic of China,3 Division of Microbial Toxins, Robert Koch Institute, D-13353 Berlin, Germany4

Received 15 March 2004/ Returned for modification 20 April 2004/ Accepted 3 May 2004

Screening for the Escherichia coli O serotype is the traditional test for identification of E. coli clones. The O-antigen gene cluster of the E. coli O114 type strain was sequenced, and 12 open reading frames were assigned functions on the basis of homology. By screening against all 186 E. coli and Shigella O serotypes, five genes specific to E. coli O114 were identified. A PCR assay based on the O-antigen-specific genes was developed and tested on 41 clinical isolates of E. coli O114. The PCR assay was shown to be highly specific and sensitive. When tested with pork and water samples, as few as 0.12 CFU of E. coli O114 g–1 were detected. Thus, the PCR assays established in this study can be used to reliably identify E. coli O114 strains and may also be used to detect E. coli O114 strains in food, water, and other environmental samples.

* Corresponding author. Mailing address: TEDA School of Biological Sciences and Biotechnology, TEDA College, Nankai University, TEDA, Tianjin 300457, People's Republic of China. Phone: 86-22-66229592. Fax: 86-22-66229596. E-mail: wanglei{at}nankai.edu.cn.

Journal of Clinical Microbiology, August 2004, p. 3799-3804, Vol. 42, No. 8
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.8.3799-3804.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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