抄録
Polymerase chain reaction (PCR) is a key technology of genetic testing, but the long thermal cycling time has been the problem to apply the rapid diagnosis of pathogenic organisms and cancer. We describe a simple, portable, and rapid genetic testing system based on ultrafast segment-flow and fluorescent detection. Especially, we improved the flow-through PCR to accelerate the thermal cycling time and simplify the operation for the practical use. The PCR solution was injected with a small volume about several microliter, and flowed as segment-flow by an air syringe. The segment of PCR solution was moved in a serpentine microchannel on a microfluidic device over two temperature zones for denaturation and annealing/extension reactions. The base plate formed upon the serpentine microchannel pattern was made of cyclo-olefin polymer (COP) and covered with a thin transparent film. Furthermore, the specialized portable system was developed for the microfluidic device of segment-flow PCR. The heater units, a syringe pump, fluorescence detector, PC were integrated in a suitcase for portable use. Using this portable system, a model microorganism of anthrax and a point mutation of FGFR3 gene were successfully detected within 6.5 min.
