Loop-mediated isothermal amplification assays for screening of bacterial integrons

文献类型： 外文期刊

第一作者： Yu, Guangchao

作者： Yu, Guangchao;Yu, Guangchao;Li, Bing;Cui, Hemiao;Chen, Siyi;Miao, Jian;Deng, Yang;Chen, Lei;Lin, Chii-wann;Bian, Huawei;Chen, Dingqiang

作者机构：

关键词： Loop-mediated isothermal amplification (LAMP);Integron screening;Bacterial integrons;Class 1 integron;Class 2 integron;Class 3 integron

期刊名称：BIOLOGICAL RESEARCH （ 影响因子：5.612； 五年影响因子：4.781 ）

ISSN： 0716-9760

年卷期： 2014 年 47 卷

页码：

收录情况： SCI

摘要： Background: The occurrence and prevalence of integrons in clinical microorganisms and their role played in antimicrobial resistance have been well studied recently. As screening and detection of integrons are concerned, current diagnostic methodologies are restricted by significant drawbacks and novel methods are required for integrons detection. Results: In this study, three loop-mediated isothermal amplification (LAMP) assays targeting on class 1, 2 and 3 integrons were implemented and evaluated. Optimization of these detection assays were performed, including studing on the reaction temperature, volume, time, sensitivity and specificity (both primers and targets). Application of the established LAMP assays were further verified on a total of 1082 isolates (previously identified to be 397 integron-positive and 685 integron-negative strains). According to the results, the indispensability of each primer had been confirmed and the optimal reaction temperature, volume and time were found to be 65 degrees C, 45 min and 25 mu L, respectively. As application was concerned, 361, 28 and 8 isolates carrying intl1, intl2 and intl3 yielded positive amplicons, respectively. Other 685 integron-negative bacteria were negative for the integron-screening LAMP assays, totaling the detection rate and specificity to be 100%. Conclusions: The intl1-, intl2- and intl3-LAMP assays established in this study were demonstrated to be the valid and rapid detection methodologies for the screening of bacterial integrons.

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