SERS Based Lateral Flow Assay for Rapid and Ultrasensitive Quantification of Dual Laryngeal Squamous Cell Carcinoma-Related miRNA Biomarkers in Human Serum Using Pd-Au Core-Shell Nanorods and Catalytic Hairpin Assembly

Abstract

Non-invasive early diagnosis is of great significant in disease pathologic development and subsequent medical treatments, and microRNA (miRNA) detection has attracted critical attention in early cancer screening and diagnosis. However, it was still a challenge to report an accurate and sensitive method for the detection of miRNA during cancer development, especially in the presence of its analogs that produce intense background noise. Herein, we developed a surface-enhanced Raman scattering (SERS)–based lateral flow assay (LFA) biosensor, assisted with catalytic hairpin assembly (CHA) amplification strategy, for the dynamic monitoring of miR-106b and miR-196b, associated with laryngeal squamous cell carcinoma (LSCC). In the presence of target miRNAs, two hairpin DNAs could self-assemble into double-stranded DNA, exposing the biotin molecules modified on the surface of palladium (Pd)–gold (Au) core–shell nanorods (Pd-AuNRs). Then, the biotin molecules could be captured by the streptavidin (SA), which was fixed on the test lines (T1 line and T2 line) beforehand. The core–shell spatial structures and aggregation Pd-AuNRs generated abundant active “hot spots” on the T line, significantly amplifying the SERS signals. Using this strategy, the limits of detections were low to aM level, and the selectivity, reproducibility, and uniformity of the proposed SERS-LFA biosensor were satisfactory. Finally, this rapid analysis strategy was successfully applied to quantitatively detect the target miRNAs in clinical serum obtained from healthy subjects and patients with LSCC at different stages. The results were consistent with the quantitative real-time PCR (qRT-PCR). Thus, the CHA-assisted SERS-LFA biosensor would become a promising alternative tool for miRNAs detection, which showed a tremendous clinical application prospect in diagnosing LSCC.

Point‐of‐Care Testing for the Detection of MicroRNAs: Towards Liquid Biopsy on a Chip

“…[78,79] Among others, those based on electrochemical and/or optical sensing platforms coupled with suitably designed microfluidics seem to be very promising to this end. Biosensing platforms relying on screen-printed electrodes, field effect transistor (FET), [80,81] surface plasmon resonance (SPR) [75,82,83] or surface-enhanced Raman scattering (SERS) [84][85][86][87] may offer the advantage of an unprecedented high sensitivity and fast detection times. Moreover, they can also be integrated with a second "dummy" sensing platform as the reference to provide a more accurate differential measurement.…”

Point‐of‐Care Testing for the Detection of MicroRNAs: Towards Liquid Biopsy on a Chip

“…[78,79] Among others, those based on electrochemical and/or optical sensing platforms coupled with suitably designed microfluidics seem to be very promising to this end. Biosensing platforms relying on screen-printed electrodes, field effect transistor (FET), [80,81] surface plasmon resonance (SPR) [75,82,83] or surface-enhanced Raman scattering (SERS) [84][85][86][87] may offer the advantage of an unprecedented high sensitivity and fast detection times. Moreover, they can also be integrated with a second "dummy" sensing platform as the reference to provide a more accurate differential measurement.…”

Lateral Flow Assay: A Summary of Recent Progress for Improving Assay Performance

“…However, they often have limited detection sensitivity when low concentrations of analyte need to be detected, so it is necessary to establish new label design procedures with high sensitivity and low cost. An alternative procedure for augmenting LFAs sensitivity is to utilize detection labels with higher absorbances that produce a stronger contrast with the background signal [ 34 , 35 , 36 , 37 ]. Song et al developed a surface-enhanced Raman scattering (SERS)-based LFA for cardiac troponin I measurement by conducting three optimization procedures.…”