Article ID Journal Published Year Pages File Type
5673947 Microbial Pathogenesis 2017 5 Pages PDF
Abstract

•Six valid and rapid LAMP assays targeting on integron genes were developed.•Gene dfrA12, orfF, aadA2, dfrA17, aadA5 and blaVIM2 were used as target genes.•272 positive and 685 negative isolates were applied on LAMP with 100% detection rate and specificity.

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 45 min and volume as 25 μl. 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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