isothiafludine and Escherichia-coli-Infections

To evaluate existing LAMP assays for detection of the seven STEC serogroups, if necessary, to improve these assays and to promote their practical application.. Pure DNA extract from 23 strains reserved in our lab was used to evaluate the existing LAMP assays. The existing LAMP assays were modified via adding 1% tetramethylene sulfoxide and 5% dimethyl sulfoxide as well as optimizing reaction temperature.. The detection limit of the modified LAMP assays was 0.1-1 pg per reaction, equivalent to 25-250 cfu per reaction, the non-specific amplification can completely be eliminated with optimal amplifying temperature, and the modified LAMP assays can specifically and sensitivly amplify targeted O serogroup-specific gene (wzx or wzy) of any concerned. In conclusion, the LAMP assays were highly susceptible to non-specific amplification caused by primer dimers, and these modified methods were free of non-specific amplification and can rapidly and reliably detect the seven major Shiga toxin-producing

Topics: Bacteriological Techniques; DNA, Bacterial; Escherichia coli; Escherichia coli Infections; Escherichia coli Proteins; Feces; Genes, Bacterial; Humans; Nucleic Acid Amplification Techniques; O Antigens; Real-Time Polymerase Chain Reaction; Sensitivity and Specificity; Serogroup; Thiazoles