School of Engineering, University of California, Santa Cruz, 1156 High Street, Santa Cruz, CA 95064, USA.
Lab Chip. 2020 Oct 21;20(20):3763-3771. doi: 10.1039/d0lc00640h. Epub 2020 Sep 7.
Bacterial pathogens resistant to antibiotics have become a serious health threat. Those species which have developed resistance against multiple drugs such as the carbapenems, are more lethal as these are last line therapy antibiotics. Current diagnostic tests for these resistance traits are based on singleplex target amplification techniques which can be time consuming and prone to errors. Here, we demonstrate a chip based optofluidic system with single molecule sensitivity for amplification-free, multiplexed detection of plasmids with genes corresponding to antibiotic resistance, within one hour. Rotating disks and microfluidic chips with functionalized polymer monoliths provided the upstream sample preparation steps to selectively extract these plasmids from blood spiked with E. coli DH5α cells. Waveguide-based spatial multiplexing using a multi-mode interference waveguide on an optofluidic chip was used for parallel detection of three different carbapenem resistance genes. These results point the way towards rapid, amplification-free, multiplex analysis of antibiotic-resistant pathogens.
对抗生素具有耐药性的细菌病原体已成为严重的健康威胁。那些对多种药物（如碳青霉烯类）产生耐药性的物种更具致命性，因为这些是最后一线治疗抗生素。目前针对这些耐药性特征的诊断测试基于单重靶标扩增技术，这种技术既耗时又容易出错。在这里，我们展示了一种基于芯片的光流控系统，该系统具有单分子灵敏度，可在一小时内对携带抗生素耐药性基因的质粒进行无扩增、多重检测。带有功能化聚合物整体柱的旋转磁盘和微流控芯片提供了上游样品制备步骤，可从用 E. coli DH5α 细胞污染的血液中选择性提取这些质粒。使用多模干涉波导在光流控芯片上进行基于波导的空间复用，用于平行检测三种不同的碳青霉烯类耐药基因。这些结果为快速、无扩增、多重分析抗生素耐药性病原体指明了方向。
