Identification of the O-antigen biosynthesis genes of Escherichia coli O91 and development of a O91 PCR serotyping test

• Published in: Journal of applied microbiology Vol. 93; no. 5; pp. 758 - 764
• Main Authors: Perelle, S, Dilasser, F, Grout, J, Fach, P
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Science Ltd 2002; Blackwell Science; Oxford University Press
• Subjects: Bacteriological methods and techniques used in bacteriology; Bacteriology; Biological and medical sciences; biosynthesis; Cloning, Molecular; DNA libraries; DNA Probes; Escherichia coli - genetics; Fundamental and applied biological sciences. Psychology; loci; Microbiology; Models, Genetic; Multigene Family; O Antigens - biosynthesis; O Antigens - genetics; O Antigens - isolation & purification; polymerase chain reaction; Polymerase Chain Reaction - methods; rapid methods; Sequence Analysis, DNA; Serotyping - methods; Shiga Toxin - genetics; Shiga toxin-producing Escherichia coli; Polymerase chain reaction; O antigen; Gene; Escherichia coli; Bacteria; Biosynthesis; Identification; Serotyping; Enterobacteriaceae
• ISSN: 1364-5072; 1365-2672
• DOI: 10.1046/j.1365-2672.2002.01743.x

Aims: The aims of the study were to characterize the O91 O-antigen gene cluster from Shiga toxin-producing Escherichia coli (STEC) O91 and to provide the basis for, a specific PCR test for rapid detection of E. coli O91. Methods and Results: The published primers complementary to JUMPstart and gnd gene, the conserved flanking sequences of O-antigen genes clusters in E. coli and related species were used to amplify the 10-kbp O91 O-antigen biosynthesis locus of STEC O91. A DNA library representative of this cluster allowed two O91 specific probes to be identified, and two specific PCR O91 serotyping tests to be successfully developed. Conclusions: These results confirm that the O-antigen gene cluster sequences of E. coli allow rapidly a specific O-antigen PCR assay to be designed. Significance and Impact of the Study: These findings increase the number of PCR-assays available to replace the classical O-serotyping among E. coli O-antigen.