Identification of Shigella serotypes by restriction of amplified O-antigen gene cluster.
Due to the scarcity of distinctive biochemical reactions for differentiation of Shigella-Escherichia coli, antigenic analysis has long been used for identification and typing of Shigella isolates. Nevertheless, several intra- and interspecific cross-reactions have been reported to disturb serotyping assays. Shigella serotyping is also occasionally affected by the transition from the smooth (S) form to the rough (R) form. Thus, there is a need for the development of novel robust and discriminating methods for Shigella identification and typing. Characteristically, all genes specifically involved in O-antigen synthesis are clustered in E. coli, Shigella, and Salmonella. Published oligonucleotide sequences complementary to JUMPstart and gene gnd, the conserved flanking sequences upstream and downstream of O-antigen gene clusters, were used to amplify the O-antigen gene cluster of representative strains of each Shigella serotype. A unique, amplified fragment was generally observed for each serotype (size ranging from 6 kbp to 17 kbp). Clearly identifiable and reproducible patterns were obtained for each serotype after MboII digestion of the products, except for S. boydii 12 which showed two distinct patterns, and S. flexneri serotypes 1 to 5 and X and Y which showed a single pattern. A database was built with the Taxotron package allowing automated identification of clinical Shigella isolates to all known serotypes.
Coimbra
R S
RS
Unité des entérobactéries, Inserm 389, Institut Pasteur, Paris, France.
Grimont
F
F
Grimont
P A
PA
D001426:Bacterial Proteins / Q000235:genetics*; D004269:DNA, Bacterial / Q000032:analysis; D015252:Deoxyribonucleases, Type II Site-Specific; D006801:Humans; D019081:O Antigens / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012703:Serotyping; D012760:Shigella / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D019081:O Antigens; C079397:rfb protein, Bacteria; C024413:endodeoxyribonuclease MboII; D015252:Deoxyribonucleases, Type II Site-Specific