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Enterotoxigenic Escherichia coli (ETEC) is the main bacterial cause of dehydrating infant diarrhoea in less-developed countries. Labile toxin (LT) is the major virulent factor of ETEC. Easy diagnostic tests are necessary to reduce the number of cases. Immunological methods have some drawbacks and also have important limitations. For that reason, a Liquid Chromatography coupled to UV detector technique (LC-UV) has been optimize to a rapid identification and quantification of LT from bacteria cultures. It is also important to know optimal conditions for LT and with this purpose several enterotoxigenic E. coli strains have been studied to determine the influence of glucose concentration and different culture media on LT production.LC-UV technique demonstrated to be a good method for LT quantification showing values of 15 ng/mL and 45 ng/mL for limits of detection and quantification respectively. LT quantification revealed that toxin production is directly related to the concentration of glucose added in the broth medium. Tryptic Soy Broth is the most efficient culture medium for E. coli growth and enterotoxin production.LC-UV method has been applied successfully in bacteria cultures to quantify LT.LT production can vary among ETEC strains and depending on the culture conditions.TSB is the most efficient culture medium for E. coli growth and toxin production.Production of LT is directly related to the presence of glucose in the medium.