Ultrafast and label-free microfluidic nucleic acid detection based on hyperspectral interferometry

Nucleic acid detection is widely used in life science and clinical medical diagnosis. Innovative methods and platform research to improve its key performances are of great significance to ensure population health, promote precision medical technology, and even ensure social stability and development. Most of the existing nucleic acid detection technologies utilized PCR as the amplification method, relying on professional and complex scientific instruments and thus is time-consuming and laborious. Fortunately, RPA offers a feasible alternative. It has the advantages of fast amplification speed, high sensitivity, simple primer design, no temperature cycle control and complex manual operations. However, the detection of amplified products is difficult and costly, and there is a lack of low-cost real-time detection methods with parallel multiple detection abilities. In this work, a label-free and real-time RPA amplicon detection method based on hyperspectral interferometry is presented. A solid-phase biochip helps to capture the RPA product in a real-time meaner and the interference spectrum signal is used to read the solid thickness increment brought by the amplicon. A Fourier domain thickness computation method contributes to calculating the thickness increase and excluding scattering noise. The detection sensitivity reaches 6 copies/reaction and the consuming time is less than 20 min. Moreover, the detection method can also be used for single point mutation readout with the specificity of merelya1%mutation-wild type ratio. Combined with a microfluidic platform, parallel, simultaneous and multiple tests can be realized with 3 microliters.