Reciprocating flow-assisted nucleic acid purification using a finger-actuated microfluidic device

Abstract

Molecular diagnostics can provide a powerful diagnostic tool since it can detect pathogens with high sensitivity, but complicated sample preparation procedures limit its widespread use as an on-site detection tool that relies on the skilled person and external equipment. To resolve these limitations, we report a solid-phase nucleic acid purification using a finger-actuated microfluidic device, which can control a set amount of flow regardless of differences in end-users. To increase the recovery rate, a finger-actuated reciprocator was newly developed and integrated into the microfluidic device that can efficiently react with silica microbeads and reagents. After verifying the finger-actuated microfluidic reciprocator, the effect of the reciprocating flow on the recovery rate was assessed to purify the standard DNA of the hepatitis B virus (HBV). The recovery rate was increased up to ∼50% and 955 to 955 000 IU mL⁻¹ of HBV standard DNA was successfully purified and detected by a real-time polymerase chain reaction. Furthermore, the proposed microfluidic device was exploited to purify the HBV DNA from the patient's blood plasma samples.