Abstract
In this study, a number of frequently detected gene cassettes from bacterial integrons have been detected and characterized by rapid and simple loop-mediated isothermal amplification (LAMP) assays. Six gene cassettes commonly found in class 1 integrons were studied, including dfrA12, dfrA17, aadA2, aadA5, orfF, and blaVIM2. Primers design, sensitivity, specificity, optimization of each LAMP assay, as well as application of the developed 6 individual LAMP assays on a large scale of bacteria, had been conducted. The optimal amplification was obtained with temperature as 65°C, reaction time span as 45min and volume as 25μl. For application, 272 isolates with various gene cassettes yielded expectable positive amplicons and other 685 integron-negative bacteria showed negative results for the LAMP assays, totaling 100% detection rate and specificity.
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