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Detection of Escherichia coli serogroup O103 by real-time polymerase chain reaction
S Perelle1, F Dilasser, J Grout
1Agence Française de Sécurité Sanitaire des Aliments, Laboratoire d'Etudes et de Recherches sur la Qualité des Aliments et sur les Procédés Agroalimentaires, Unité EBA: Etude moléculaire des contaminants biologiques alimentaires, Maisons-Alfort, France.
Aims:
The aims of the study were to identify the specific genes of O-antigen gene cluster from Shiga toxin-producing Escherichia coli (STEC) O103 and to provide the basis for a specific real-time PCR test for rapid detection of E. coli O103.
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 12-kbp O103 O-antigen biosynthesis locus of STEC O103. A DNA library representative of this cluster allowed two O103-specific probes to be identified in the flippase (wzx) and UDP-galactose-4-epimerase (galE) genes. Two specific O103 serotyping real-time PCR tests based on these two genes were successfully developed.
Conclusions:
These results confirm that the O-antigen gene cluster sequences of E. coli allow rapidly a specific O-antigen real-time PCR assay to be designed.
Significance And Impact Of The Study:
These findings increase the number of real-time PCR-assays available to replace the classical O-serotyping among E. coli O-antigen.
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