Abstract
Since nucleic acid amplification technology has become a vital tool for disease diagnosis, the development of precise applied nucleic acid detection technologies in point-of care testing (POCT) has become more significant. The microfluidic-based nucleic acid detection platform offers a great opportunity for on-site diagnosis efficiency, and the system is aimed at user-friendly access. Herein, we demonstrate a microfluidic system with simple operation that provides reliable nucleic acid results from 18 uniform droplets via LAMP detection. By using only micropipette regulation, users are able to control the nanoliter scale of the droplets in this valve-free and pump-free microfluidic (MF) chip. Based on the oil enclosure method and impermeable fabrication, we successfully preserved the reagent inside the microfluidic system, which significantly reduced the fluid loss and condensation. The relative standard deviation (RSD) of the fluorescence intensity between the droplets and during the heating process was < 5% and 2.0%, respectively. Additionally, for different nucleic acid detection methods, the MF-LAMP chip in this study showed good applicability to both genome detection and gene expression analysis.
Highlights
Since nucleic acid amplification technology has become a vital tool for disease diagnosis, the development of precise applied nucleic acid detection technologies in point-of care testing (POCT) has become more significant
An elimination of the temperature steps in traditional polymerase chain reaction (PCR) by circumventing the thermal heating and cooling system. This can be achieve by integrated the system with isothermal nucleic acid amplification methods, like loop-mediated isothermal amplification (LAMP), that is available for amplifying nucleic acid at a single temperature
We demonstrate a simple operational LAMP assay that contains uniform droplets, each of which controlled by a capillary channel for the fluid exchange and oil enclosed
Summary
Since nucleic acid amplification technology has become a vital tool for disease diagnosis, the development of precise applied nucleic acid detection technologies in point-of care testing (POCT) has become more significant. The flow in the microfluidic is typically drive and regulate by capillary force on the chip providing robust and easy to use advantages without any active components Such an emerging strategy for the movement of nucleic acid samples is achieved with characteristics including easy fabrication, a lack of external power, low cost, compactness and portability. The sealed techniques including block inlet and outlet with tape or PDMS by O 2 plasma treatment bonding; close the air channel by vacuum system in attempt to avoid reagent condense[37,38,39,40,41] To address their easy integration of such devices, the use of a passive-driven pumping system without further additional instruments is favourable as a working principle for all users
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