A portable, shock-proof, surface-heated droplet PCR system for Escherichia coli detection.
A portable, shock-proof, surface-heated droplet PCR system for Escherichia coli detection.
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DOI：
10.1016/j.bios.2015.06.026
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发表时间：
2015-12-15
影响因子：
12.6
通讯作者：
Yoon JY
中科院分区：
文献类型：
--
作者：
Angus SV;Cho S;Harshman DK;Song JY;Yoon JY
开发了一种新型聚合酶链反应（PCR）装置，该装置使用导线引导液滴操作（WDM）来引导液滴通过三个不同的加热室。PCR扩增后，使用基于智能手机的荧光显微镜进行终点检测。该装置用于鉴定大肠杆菌基因组DNA中16S rRNA基因V3高变区。检测下限为每个样本103个基因组拷贝。该设备是便携式的，具有基于智能手机的终点检测功能，可以快速准确地提供分析结果（30个周期扩增15分钟）。由于液滴与热电偶和加热器表面的聚四氟乙烯薄膜之间有多个接触点，该系统还具有抗冲击和抗振动性能。热电偶还提供实时液滴温度反馈，以确保在移动到PCR的下一个腔室/步骤之前达到设定的温度。该设备配备使用硅油或椰子油。椰子油提供了额外的便携性和易于运输，因为它的高熔化温度意味着它在室温下是固体。
A novel polymerase chain reaction (PCR) device was developed that uses wire-guided droplet manipulation (WDM) to guide a droplet over three different heating chambers. After PCR amplification, end-point detection is achieved using a smartphone-based fluorescence microscope. The device was tested for identification of the 16S rRNA gene V3 hypervariable region from Escherichia coli genomic DNA. The lower limit of detection was 103 genome copies per sample. The device is portable with smartphone-based end-point detection and provides the assay results quickly (15 min for a 30-cycle amplification) and accurately. The system is also shock and vibration resistant, due to the multiple points of contact between the droplet and the thermocouple and the Teflon film on the heater surfaces. The thermocouple also provides realtime droplet temperature feedback to ensure it reaches the set temperature before moving to the next chamber/step in PCR. The device is equipped to use either silicone oil or coconut oil. Coconut oil provides additional portability and ease of transportation by eliminating spilling because its high melting temperature means it is solid at room temperature.