摘要

抗微生物耐药细菌的迅速出现需要开发新的诊断测试。通过检测编码该抗性的基因，基于核酸的测定确定抗微生物药物敏感性。在这项研究中，我们证明快速和同时检测三种基因，赋予细菌对广谱β-内酰胺和碳青霉烯类抗生素的耐药性; CTX-M-15，KPC和NDM-1。该测定使用在可编程数字微流体（DMF）平台上实施的等温DNA扩增（重组酶聚合酶扩增，RPA）。 使用自动分配方案同时操作45个含有样品DNA，试剂和对照的nL体积液滴。在DMF装置上电子控制下处理和混合液滴，通过荧光测量正扩增。这些设备上的测定法在台式测定时间到达时间（TTP）的一半时显着改善。

The rapid emergence of antimicrobial resistant bacteria requires the development of new diagnostic tests. Nucleic acid-based assays determine antimicrobial susceptibility by detecting genes that encode for the resistance. In this study, we demonstrate rapid and simultaneous detection of three genes that confer resistance in bacteria to extended spectrum β-lactam and carbapenem antibiotics; CTX-M-15, KPC and NDM-1. The assay uses isothermal DNA amplification (recombinase polymerase amplification, RPA) implemented on a programmable digital microfluidics (DMF) platform. Automated dispensing protocols are used to simultaneously manipulate 45 droplets of nL volume containing sample DNA, reagents, and controls. The droplets are processed and mixed under electronic control on the DMF devices with positive amplification measured by fluorescence. The assay on these devices is significantly improved with a Time to Positivity (TTP) half that of the benchtop assay.

http://www.mdpi.com/2072-666X/8/4/111