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Alkaline Phosphatase-conjugated Oligonucleotide Probes For Enterotoxigenic Escherichia Coli In Travelers To South America And West Africa

J J Oprandy, S A Thornton, C H Gardiner, D Burr, R Batchelor, A L Bourgeois

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Several studies have demonstrated the usefulness of 32P-labeled recombinant DNA probes for identifying enterotoxigenic Escherichia coli (ETEC). The use of radioisotopes and X-ray development, however, severely handicaps the utility of DNA probes in most clinical laboratories. In this study, enzyme-labeled oligonucleotide probes for ETEC LT (heat-labile toxin) and ST (heat-stable toxin) genes were compared with the standard Y1 adrenal cell and suckling mouse assays for their ability to identify ETEC in a population of American adults experiencing acute episodes of diarrhea in South America and West Africa. The LT probe hybridized with 12% (64 of 529) of the E. coli colonies tested, whereas 11% (57 of 529) were positive by Y1 adrenal cell assay. DNA from 9% (47 of 529) of the E. coli colonies tested hybridized with the ST probe, whereas only 5% (28 of 529) produced ST as measured by the suckling mouse bioassay. For the patient samples tested, correlation between probe and bioassay for LT was 97%, or three discrepancies in 111 patients tested. Overall concordance of the ST probe and bioassay was 95%, or five discrepancies in 111 patients. Enzyme-labeled oligonucleotide probes represent a major advance in the diagnosis of ETEC-associated diarrheal disease and may be used in laboratories with minimal equipment.