KIST Europe , Campus E7.1, 66123 Saarbrücken, Germany.
Northwestern Polytechnical University (NPU) , School of Mechanical Engineering, Department of Microsystem Engineering, 127 West Youyi Road, Xi'an, Shaanxi 710072, P. R. China.
Anal Chem. 2016 May 3;88(9):4803-7. doi: 10.1021/acs.analchem.6b00278. Epub 2016 Apr 21.
We show the utilization of a recently developed cellphone-sized real-time polymerase chain reaction (PCR) device to detect Ebola virus RNA using single-step reverse transcription PCR (RT-PCR). The device was shown to concurrently perform four PCRs, each with a sample volume of 100 nL: one positive control with both Ebola and GAPDH RNA and one negative control. The last two positions were used to measure the GAPDH and the Ebola content of a sample. A comparison of threshold cycles (CT) from the two samples provided relative quantification. The entire process, which consisted of reverse transcription, PCR amplification, and melting curve analysis (MCA), was conducted in less than 37 min. The next step will be integration with a sample preparation unit to form an integrated sample-to-answer system for point-of-care infectious disease diagnostics.
我们展示了一种最近开发的手机大小的实时聚合酶链反应(PCR)设备,该设备可用于使用一步法逆转录 PCR(RT-PCR)检测埃博拉病毒 RNA。该设备可同时进行四个 PCR 反应,每个反应的样本体积为 100nL:一个含有埃博拉病毒和 GAPDH RNA 的阳性对照以及一个阴性对照。最后两个位置用于测量样本中的 GAPDH 和埃博拉病毒含量。从两个样本的阈值循环(CT)进行比较,可提供相对定量。整个过程包括逆转录、PCR 扩增和熔解曲线分析(MCA),耗时不到 37 分钟。下一步将与样品制备单元集成,形成用于即时传染病诊断的集成样品到答案系统。