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3× multiplexed detection of antibiotic resistant plasmids with single molecule sensitivity
Lab on a Chip ( IF 6.1 ) Pub Date : 2020-09-02 , DOI: 10.1039/d0lc00640h G G Meena 1 , R L Hanson , R L Wood , O T Brown , M A Stott , R A Robison , W G Pitt , A T Woolley , A R Hawkins , H Schmidt
Lab on a Chip ( IF 6.1 ) Pub Date : 2020-09-02 , DOI: 10.1039/d0lc00640h G G Meena 1 , R L Hanson , R L Wood , O T Brown , M A Stott , R A Robison , W G Pitt , A T Woolley , A R Hawkins , H Schmidt
Affiliation
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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.
中文翻译：
具有单分子灵敏度的抗生素抗性质粒的 3× 多重检测
对抗生素具有抗药性的细菌病原体已成为严重的健康威胁。那些对碳青霉烯类等多种药物产生耐药性的物种更具致命性，因为它们是最后一线治疗抗生素。目前针对这些抗性特征的诊断测试基于单重靶标扩增技术，该技术可能非常耗时且容易出错。在这里，我们展示了一种基于芯片的光流控系统，具有单分子灵敏度，可在一小时内对带有抗生素耐药性相关基因的质粒进行无扩增、多重检测。带有功能化聚合物整体的旋转盘和微流控芯片提供了上游样品制备步骤，以选择性地从掺有大肠杆菌DH5α 细胞的血液中提取这些质粒。使用光流控芯片上的多模干涉波导的基于波导的空间多路复用用于并行检测三种不同的碳青霉烯类抗性基因。这些结果为对抗生素耐药病原体进行快速、无扩增、多重分析指明了道路。
更新日期：2020-10-13
中文翻译：
具有单分子灵敏度的抗生素抗性质粒的 3× 多重检测
对抗生素具有抗药性的细菌病原体已成为严重的健康威胁。那些对碳青霉烯类等多种药物产生耐药性的物种更具致命性，因为它们是最后一线治疗抗生素。目前针对这些抗性特征的诊断测试基于单重靶标扩增技术，该技术可能非常耗时且容易出错。在这里，我们展示了一种基于芯片的光流控系统，具有单分子灵敏度，可在一小时内对带有抗生素耐药性相关基因的质粒进行无扩增、多重检测。带有功能化聚合物整体的旋转盘和微流控芯片提供了上游样品制备步骤，以选择性地从掺有大肠杆菌DH5α 细胞的血液中提取这些质粒。使用光流控芯片上的多模干涉波导的基于波导的空间多路复用用于并行检测三种不同的碳青霉烯类抗性基因。这些结果为对抗生素耐药病原体进行快速、无扩增、多重分析指明了道路。
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