Cao Weidong, Bean Brian, Corey Scott, Coursey Johnathan S, Hasson Kenton C, Inoue Hiroshi, Isano Taisuke, Kanderian Sami, Lane Ben, Liang Hongye, Murphy Brian, Owen Greg, Shinoda Nobuhiko, Zeng Shulin, Knight Ivor T
Canon U.S. Life Sciences, Inc, Rockville, MD, USA.
Sobosoft LLC, Catonsville, MD, USA.
J Lab Autom. 2016 Jun;21(3):402-11. doi: 10.1177/2211068215579015. Epub 2015 Mar 31.
We report the development of an automated genetic analyzer for human sample testing based on microfluidic rapid polymerase chain reaction (PCR) with high-resolution melting analysis (HRMA). The integrated DNA microfluidic cartridge was used on a platform designed with a robotic pipettor system that works by sequentially picking up different test solutions from a 384-well plate, mixing them in the tips, and delivering mixed fluids to the DNA cartridge. A novel image feedback flow control system based on a Canon 5D Mark II digital camera was developed for controlling fluid movement through a complex microfluidic branching network without the use of valves. The same camera was used for measuring the high-resolution melt curve of DNA amplicons that were generated in the microfluidic chip. Owing to fast heating and cooling as well as sensitive temperature measurement in the microfluidic channels, the time frame for PCR and HRMA was dramatically reduced from hours to minutes. Preliminary testing results demonstrated that rapid serial PCR and HRMA are possible while still achieving high data quality that is suitable for human sample testing.
我们报告了一种基于微流控快速聚合酶链反应（PCR）和高分辨率熔解分析（HRMA）的用于人类样本检测的自动化基因分析仪的开发情况。集成的DNA微流控芯片盒被用于一个配备机器人移液器系统的平台上，该系统通过依次从384孔板中吸取不同的测试溶液，在移液器吸头中混合，然后将混合液输送到DNA芯片盒来工作。开发了一种基于佳能5D Mark II数码相机的新型图像反馈流量控制系统，用于在不使用阀门的情况下控制流体通过复杂的微流控分支网络的流动。同一台相机用于测量在微流控芯片中产生的DNA扩增子的高分辨率熔解曲线。由于微流控通道中快速的加热和冷却以及灵敏的温度测量，PCR和HRMA的时间框架从数小时大幅缩短至数分钟。初步测试结果表明，快速连续PCR和HRMA是可行的，同时仍能获得适用于人类样本检测的高质量数据。
