Miniaturized systems with microchips for four isothermal amplification reactions

Abstract

A real-time fluorescence detection biomedical system with a microfluidic chip was developed for loop-mediated isothermal amplification (LAMP) reactions. Two kinds of microfluidic chips with linear microchannels or microcells were designed and fabricated by MEMS technology and thin-casting method. Both traditional centrifugal tubes and microfluidic chips were used for LAMP reactions. Based on the experimental results of end-point fluorescence detection, the microfluidic chip can be successfully used for LAMP reactions. And then our real-time fluorescence detection biomedical system with a microfluidic chip was used to implement positive reaction and negative control. The volume of the LAMP reaction was greatly reduced from about 25 μL to 2 μL~3 μL and the reaction time can be reduced from one hour to half an hour by using our real-time fluorescence detection biomedical system. This biomedical system has the potential for point-of-care diagnostics (POCT) with the many advantages, such as not high cost, short analysis and detection time, not much reagent and sample consumption and so on.