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Journal of Clinical Microbiology, October 2005, p. 5143-5149, Vol. 43, No. 10
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.10.5143-5149.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Development of PCR Assays Targeting the Genes Involved in Synthesis and Assembly of the New Escherichia coli O174 and O177 O Antigens

Lothar Beutin,1,2 Qingke Kong,3,4 Lu Feng,3,4 Quan Wang,3,4 Gladys Krause,2,5 Luciana Leomil,1,6 Qi Jin,7 and Lei Wang3,4*

Division of Microbial Toxins, Robert Koch Institut, Nordufer 20, D-13353 Berlin, Germany,1 National Reference Laboratory for Escherichia coli, Federal Institute for Risk Assessment (BfR), D-12277 Berlin, Germany,2 TEDA School of Biological Sciences and Biotechnology, Nankai University, 23 Hong Da Street, TEDA, Tianjin 300457, Peoples Republic of China,3 Tianjin Key Laboratory for Microbial Functional Genomics, TEDA College, Nankai University, 23 Hong Da Street, TEDA, Tianjin 300457, Peoples Republic of China,4 Freie Universität Berlin, Königin Luise Str. 12-16, D-14195 Berlin, Germany,5 Departamento de Microbiologia, Instituto de Ciências Biomédicas II, Universidade de São Paulo, 05508-900 São Paulo, Brazil,6 State Key Laboratory for Molecular Virology and Genetic Engineering, Beijing 100052, Peoples Republic of China7

Received 16 April 2005/ Returned for modification 13 June 2005/ Accepted 11 July 2005

Escherichia coli O174 and O177 are newly described O serogroups which were reported as human pathogens. Identification of these strains by serotyping has been restricted, as the required sera are not commercially available. In this study, a collection of 13 E. coli O174 strains and 12 E. coli O177 strains was studied on the O:H serotypes and virulence markers. The O-antigen gene clusters of E. coli O174 and O177 were sequenced, and associated genes were assigned functions on the basis of homology. Two genes, each specific for E. coli O174 and O177, were identified. PCR assays based on the O-antigen-specific genes were developed and tested on 25 clinical and environmental isolates of those two serogroups as well as 26 isolates of other O serogroups. As little as 1 pg per µl of chromosomal DNA and as few as 0.1 CFU per g of pork and water samples were detected for either strain. The PCR assays established in this study were shown to be highly sensitive and reliable and could be the method of choice for detection of these two human pathogens from clinical, food, and other environmental samples.

* Corresponding author. Mailing address: Tianjin Key Laboratory for Microbial Functional Genomics, TEDA College, Nankai University, 23 Hong Da Street, TEDA, Tianjin 300457, Peoples Republic of China. Phone: 86-22-66229588. Fax: 86-22-66229596. E-mail: wanglei{at}nankai.edu.cn.

Journal of Clinical Microbiology, October 2005, p. 5143-5149, Vol. 43, No. 10
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.10.5143-5149.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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