Comparison of polymerase chain reaction systems for detection of different cdt genes in Escherichia coli strains

Letters in Applied Microbiology
Mana Oloomi, Saeid Bouzari

Abstract

Cytolethal distending toxins (CDT) are tripartite toxins encoded by three adjacent or overlapping genes (cdtA, cdtB, cdtC) and found in multiple pathogens. The present knowledge regarding heterogeneity of cdt genes and our previous study revealed that the available polymerase chain reaction (PCR) systems lack adequate specificity. The detection of various cdt genes present in Escherichia coli strains, from different geographical regions demands further assays for wide-range coverage. On the basis of these observations, we were prompted to undertake the present study; hence the specificity of existing PCR systems was addressed using E. coli prototype strains with known cdt gene sequences. A multiplex PCR designed for the detection of E. coli cdt genes was found to be sensitive and specific enough for initial screening. However, for subtyping, the PCR systems yielded nonspecific products upon amplification. These primers are usually designed for sequences of the cdtB locus (the most conserved region of the gene), and since CDT-producing E. coli strains carry different cdt genes, none of the systems are really type specific. Furthermore, PCR systems with type-specific primers for other regions of the gene, i.e. ORF A or ORF C are ...Continue Reading

References

Apr 15, 1997·Proceedings of the National Academy of Sciences of the United States of America·L D CopeE J Hansen
Sep 6, 2003·Journal of Clinical Microbiology·Sigrid Van BostJacques Mainil
Jan 27, 2004·Infection and Immunity·Carol L PickettNancy A Strockbine

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Citations

Mar 7, 2008·APMIS : Acta Pathologica, Microbiologica, Et Immunologica Scandinavica·Mana Oloomi, Saeid Bouzari

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