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Biosensors 2015, 5(1), 51-68; doi:10.3390/bios5010051

Escherichia coli O-Antigen Gene Clusters of Serogroups O62, O68, O131, O140, O142, and O163: DNA Sequences and Similarity between O62 and O68, and PCR-Based Serogrouping

1
Molecular Characterization of Foodborne Pathogens Research Unit, Eastern Regional Research Center, Agricultural Research Service, U.S. Department of Agriculture, Wyndmoor, PA 19038, USA
2
Reference Center, Department of Veterinary and Biomedical Sciences, The Pennsylvania State University, University Park, PA 16802, USA
3
Animal Genomics and Improvement Laboratory, Agricultural Research Service, U.S. Department of Agriculture, Beltsville, MD 20705, USA
4
Craig Venter Institute, Rockville, MD 20850, USA
5
Department of Nutrition & Food Science, University of Maryland, College Park, MD 20742, USA
*
Author to whom correspondence should be addressed.
Academic Editor: Glen C. Rains
Received: 9 December 2014 / Revised: 24 January 2015 / Accepted: 30 January 2015 / Published: 5 February 2015
(This article belongs to the Special Issue Biosensors in Agroecosystems)
View Full-Text   |   Download PDF [454 KB, uploaded 5 February 2015]   |  

Abstract

The DNA sequence of the O-antigen gene clusters of Escherichia coli serogroups O62, O68, O131, O140, O142, and O163 was determined, and primers based on the wzx (O-antigen flippase) and/or wzy (O-antigen polymerase) genes within the O-antigen gene clusters were designed and used in PCR assays to identify each serogroup. Specificity was tested with E. coli reference strains, field isolates belonging to the target serogroups, and non-E. coli bacteria. The PCR assays were highly specific for the respective serogroups; however, the PCR assay targeting the O62 wzx gene reacted positively with strains belonging to E. coli O68, which was determined by serotyping. Analysis of the O-antigen gene cluster sequences of serogroups O62 and O68 reference strains showed that they were 94% identical at the nucleotide level, although O62 contained an insertion sequence (IS) element located between the rmlA and rmlC genes within the O-antigen gene cluster. A PCR assay targeting the rmlA and rmlC genes flanking the IS element was used to differentiate O62 and O68 serogroups. The PCR assays developed in this study can be used for the detection and identification of E. coli O62/O68, O131, O140, O142, and O163 strains isolated from different sources. View Full-Text
Keywords: PCR; Escherichia coli; serogroups; DNA sequence; O-antigen gene cluster; detection; identification PCR; Escherichia coli; serogroups; DNA sequence; O-antigen gene cluster; detection; identification
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Liu, Y.; Yan, X.; DebRoy, C.; Fratamico, P.M.; Needleman, D.S.; Li, R.W.; Wang, W.; Losada, L.; Brinkac, L.; Radune, D.; Toro, M.; Hegde, N.; Meng, J. Escherichia coli O-Antigen Gene Clusters of Serogroups O62, O68, O131, O140, O142, and O163: DNA Sequences and Similarity between O62 and O68, and PCR-Based Serogrouping. Biosensors 2015, 5, 51-68.

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