Of late, many nucleic acid analysis
platforms have been established,
but there is still room for constructing integrated nucleic acid detection
systems with high nucleic acid extraction efficiency, low detection
cost, and convenient operation. In this work, a simple rotary valve-assisted
fluidic chip coupling with CRISPR/Cas12a was established to achieve
fully integrated nucleic acid detection. All of the detection reagents
were prestored on the fluidic chip. With the aid of the rotary valve
and syringe, the liquid flow and stirring can be precisely controlled.
The nucleic acid extraction, loop-mediated isothermal amplification
(LAMP) reaction, and CRISPR detection could be completed in 80 min.
A clean reservoir and an air reservoir on the fluidic chip were designed
to effectively remove the remaining ethanol. With Vibrio
parahaemolyticus as the targets, the detection sensitivity
of the fluidic chip could reach 3.1 × 101 copies of
target DNA per reaction. A positive sample could be sensitively detected
by CRISPR/Cas12a to produce a green fluorescent signal, while a negative
sample generated no fluorescent signal. Further, the fluidic chip
was successfully applied for detection of spiked shrimp samples, which
showed the same detection sensitivity. A great feasibility for real-sample
detection was showed by the fluidic chip. The proposed detection platform
did not need expensive centrifugal instruments or pumps, which displayed
its potential to become a powerful tool for food safety analysis and
clinical diagnostics, especially in the resource-limited areas.