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ARS Home » Northeast Area » Wyndmoor, Pennsylvania » Eastern Regional Research Center » Characterization and Interventions for Foodborne Pathogens » Research » Publications at this Location » Publication #302164
Title: DNA sequence and analysis of the O-antigen gene clusters of Escherichia coli serogroups O62, O68, O131, O140, O142, and O163 and serogroup-specific PCR assays

Author
item Liu, Yanhong
item Fratamico, Pina
item DEBROY, CHITRITA - Pennsylvania State University
item Yan, Xianghe
item Needleman, David
item Li, Robert
item WANG, WEI - J Craig Venter Institute
item LOSADA, LILIANA - J Craig Venter Institute
item BRINKAC, LAUREN - J Craig Venter Institute
item RODUNE, DIANA - J Craig Venter Institute
item TORO, MAGALY - University Of Maryland
item MENG, JIANGHONG - University Of Maryland

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 1/26/2014
Publication Date: 3/22/2014
Citation: Liu, Y., Fratamico, P.M., Debroy, C., Yan, X., Needleman, D.S., Li, R.W., Wang, W., Losada, L., Brinkac, L., Rodune, D., Toro, M., Meng, J. 2014. DNA sequence and analysis of the O-antigen gene clusters of Escherichia coli serogroups O62, O68, O131, O140, O142, and O163 and serogroup-specific PCR assays. Meeting Abstract. March 22-25, 2014.

Interpretive Summary:

Technical Abstract: The DNA sequence of the O-antigen gene clusters of Escherichia coli serogroups O62, O68, O131, O140, O142, and O163 was determined. There were 9 to 12 open reading frames (ORFs) identified, encoding genes required for O-antigen sugar biosynthesis, transfer, and processing. Primers based on the wzx (O-antigen flippase) and wzy (O-antigen polymerase) genes within the O-antigen gene clusters were designed and used in PCR assays to identify each serogroup. Of interest, an NCBI BLAST alignment showed that the O-antigen gene cluster of O62 shares a 99% sequence identity with O68, with the exception of an insertion sequence (IS) element between the rmlA and rmlC genes of O62. The IS insertion in O62 infers that O68 may be the ancestor of O62. Specificity testing using strains belonging to each of the serogroups isolated from various sources, representative standard strains of 174 E. coli O serogroups, and 16 non-E. coli bacteria revealed that the PCR assays were specific for each serogroup. The PCR assay targeting O62 also gave positive results with strains serotyped as E. coli O68, which confirms that the O-antigen gene cluster sequences for these two serogroups are very similar. 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.