Detection of Escherichia coli serogroup O103 by real-time polymerase chain reaction.
OBJECTIVE
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.


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.


CONCLUSIONS
These findings increase the number of real-time PCR-assays available to replace the classical O-serotyping among E. coli O-antigen.
Perelle
S
S
Agence 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.
Dilasser
F
F
Grout
J
J
Fach
P
P
D015139:Blotting, Southern; D015342:DNA Probes / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis*; D004926:Escherichia coli / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D004927:Escherichia coli Infections / Q000175:diagnosis* / Q000382:microbiology; D005516:Food Microbiology; D006801:Humans; D019081:O Antigens / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D012703:Serotyping; D022641:Shiga Toxin 2 / Q000096:biosynthesis
D015342:DNA Probes; D004269:DNA, Bacterial; D019081:O Antigens; D022641:Shiga Toxin 2