Rombach Markus, Hin Sebastian, Specht Mara, Johannsen Benita, Lüddecke Jan, Paust Nils, Zengerle Roland, Roux Louis, Sutcliffe Thomas, Peham Johannes R, Herz Christopher, Panning Marcus, Donoso Mantke Oliver, Mitsakakis Konstantinos
Hahn-Schickard, Georges-Koehler-Allee 103, 79110 Freiburg, Germany.
Analyst. 2020 Oct 26;145(21):7040-7047. doi: 10.1039/d0an01226b.
We present the RespiDisk enabling the fully automated and multiplex point-of-care (POC) detection of (currently) up to 19 respiratory tract infection (RTI) pathogens from a single sample based on reverse transcriptase polymerase chain reaction (RT-PCR). RespiDisk comprises a RTI-specific implementation of the centrifugal microfluidic LabDisk platform and combines new and existing advanced unit operations for liquid control, thereby automating all assay steps only by a spinning frequency and temperature protocol in combination with the use of a permanent magnet for in situ bead handing. The capabilities of the system were demonstrated with 36 tested quality samples mimicking clinical conditions (clinical and/or cultured material suspended in transport medium or synthetic bronchoalveolar lavage (BAL)) from past external quality assessment (EQA) panels covering 13 of the 19 integrated RTI detection assays. In total, 36 samples × 19 assays/sample resulting in 684 assays were performed with the RespiDisk, and its analytical performance was in full agreement with the routine clinical workflow serving as reference. A strong feature of the platform is its universality since its components allow the simultaneous detection of a broad panel of bacteria and viruses in a single run, thereby enabling the differentiation between antibiotic-treatable diseases. Furthermore, the full integration of all necessary biochemical components enables a reduction of the hands-on time from manual to automated sample-to-answer analysis to about 5 min. The study was performed on an air-heated LabDisk Player instrument with a time-to-result of 200 min.
我们展示了RespiDisk，它能够基于逆转录聚合酶链反应（RT-PCR）从单个样本中全自动、多重即时检测（POC）（目前）多达19种呼吸道感染（RTI）病原体。RespiDisk包括离心微流控LabDisk平台的RTI特异性实施方案，并结合了用于液体控制的新的和现有的先进单元操作，从而仅通过旋转频率和温度协议以及使用永久磁铁进行原位磁珠处理来自动化所有检测步骤。该系统的性能通过36个模拟临床条件的测试质量样本（悬浮在运输介质或合成支气管肺泡灌洗（BAL）中的临床和/或培养材料）得到了验证，这些样本来自过去外部质量评估（EQA）小组，涵盖了19种集成RTI检测方法中的13种。使用RespiDisk总共进行了36个样本×19种检测/样本，共684次检测，其分析性能与作为参考的常规临床工作流程完全一致。该平台的一个强大特点是其通用性，因为其组件允许在一次运行中同时检测多种细菌和病毒，从而能够区分可使用抗生素治疗的疾病。此外，所有必要生化组件的完全集成使从手动到自动的样本到答案分析的操作时间减少到约5分钟。该研究在空气加热的LabDisk Player仪器上进行，结果报告时间为200分钟。
