Rapid Detection of microRNA-122 in Serum and Finger Blood Using a Lateral Flow Nucleic Acid Biosensor.

Abstract

MicroRNA122 (miR-122) is a microRNA that is highly expressed in hepatocytes and has been identified as a prospective therapeutic target and biomarker for liver injury. An expanding body of research has demonstrated that miR-122 is a critical regulator in both the initiation and progression of a wide range of liver diseases. Traditional methods for detecting miR-122 mainly include Northern blotting and qRT-PCR, but they are technically complex and cumbersome, requiring expensive instruments and high technical requirements. In this paper, we present a novel rapid testing method utilizing a lateral flow nucleic acid biosensor (LFNAB) for the sensitive and time-efficient detection of miR-122. This approach offers several advantages, including a high specificity for miR-122, the ability to detect low concentrations of the target molecule, and a significantly reduced testing time compared to conventional detection methods. In this study, a thiol-modified single-stranded detection DNA probe (Det-DNA), a biotinylated single-stranded capture DNA probe (Cap-DNA), and a biotinylated single-stranded control DNA probe (Con-DNA) are used to construct the LFNAB. A gold nanoparticle (AuNP) is a colored tag, which is used to label the Det-DNA probe. The principle of detecting miR-122 is based on dual DNA-miRNA hybridization reactions on the LFNAB to form sandwich-type AuNP-Det-DNA-miR-122-Cap-DNA complexes, which are captured on the test area of LFNAB for visualization and quantification. After systematic optimization of conditions of experiment, the response of LFNAB was highly linear within the scope of 0 pM-100 pM miR-122, and the detection limit in 15 min was 3.90 pM. The use of LFNAB to detect miR-122 in serum and fingertip blood has yielded satisfactory results. This successful application indicates the effectiveness of LFNAB in detecting miR-122 in both serum and fingertip blood samples, showcasing its potential utility in clinical and research settings for assessing miR-122 levels in different biological samples.