Search

Cervical cancer is a common cause of cancer-related death in women worldwide, and quantitative detection of serum tumor markers has practical implications for diagnosing cervical cancer. In this study, a novel surface-enhanced Raman scattering (SERS)-based immunosensor for the hypersensitive simultaneous detection of survivin and osteopontin (OPN) in cervical cancer serum was proposed, which was composed of the AuNPs@SiO2 microspheres (Au@SiO2) array immunosubstrate and Au-Ag nanoshells (Au-AgNSs) immunoprobes. The experimental results demonstrated that the constructed SERS immunosensor exhibited satisfactory selectivity and reproducibility while possessing detection limits as low as 0.908 pg/mL and 0.813 pg/mL for survivin and OPN in human serum. Finally, its usefulness in real serum samples (from healthy subjects, cervical intraepithelial neoplasia 1 (CIN 1), CIN 2, CIN 3 and cervical cancer patients) was verified, and the detection data were in good agreement with the enzyme-linked immunosorbent assay results. This immunosensor has excellent performance, which is promising for the early screening of cancer. The ordered single-layer gold film and monolayer SiO2 array were obtained by the oil–water interface self-assembly method and Langmuir-Blodgett (LB) gas–liquid interface assembly method, respectively. Then, these two ordered structures are combined to prepare Au@SiO2 array substrates. This ordered array substrate not only ensures good uniformity, but also improves SERS performance. [ABSTRACT FROM AUTHOR]

Research methods for DNA detection have been widely extended since the application of nanotechnology, but it remains a challenge to detect specific DNA sequences or low abundance genes in biological samples with accuracy and sensitivity. Herein, a novel biosensor is proposed for the analysis of target DNA in maize based on a high-density "hot spot" surface-enhanced Raman scattering (SERS) substrate and hybridization chain reaction (HCR) amplification strategy. Finite-difference time-domain (FDTD) simulation confirmed that the electromagnetic field enhancement at the gaps between the adjacent Au nanoflowers (AuNFs) was much richer than that around a single AuNF, indicating that the aggregated AuNFs had a good SERS signal amplification effect. Based on the high-density "hot spots" AuNF substrate, the HCR of biotin-hairpin DNA (bio-H1 and bio-H2) could be triggered by the existence of target DNA and a long double-stranded DNA (dsDNA) could be formed rapidly. Due to the biotin and streptavidin (SA) interaction, SERS probes prepared by modifying biotin-DNA-cyanine 5 (bio-DNA-Cy5) on the surface of Ag–Au core–shell nanorods (Ag–AuNRs) could be assembled along the dsDNA, causing significant signal enhancement. The developed biosensor displayed a significant sensitivity with a broad detection range covering from 0.01 to 1 × 104 fM, and the limit of detection (LOD) was as low as 6.31 aM. Moreover, SERS was applied to detect target DNA in maize extract to verify the feasibility of the detection strategy. This strategy could not only carry out the ultrasensitive detection of target DNA but also provide an informative supplement to the SERS biosensing platform. [ABSTRACT FROM AUTHOR]

Circulating tumour DNAs (ctDNAs) have been reported to be associated with real-time information of progression; however, an accurate and sensitive method has not been established. Herein, a novel dual-signal amplification strategy based on a pump-free surface-enhanced Raman scattering (SERS) microfluidic chip and a catalytic hairpin assembly (CHA) technique was developed for the dynamic monitoring of BRAF V600E and KRAS G12V, which are two non-small cell lung cancer (NSCLC)-related ctDNAs. In the presence of targets, CHA reactions can be triggered between two hairpin DNAs, fixing Pd-Au core-shell nanorods (Pd-AuNRs) onto the magnetic beads (MBs) surface. Thereafter, the composite structures can assemble under the action of magnet, enabling dual-amplification of SERS signal. Using this strategy, the limit of detection (LOD) was low (i.e. at the aM level) in serum. Furthermore, the entire chip-based analysis process could be completed within 5 min, eliminating manual incubation and heavy-duty injection pumps. The selectivity, reproducibility and uniformity of the proposed pump-free SERS microfluidic chip were satisfactory. This superior analysis strategy was finally applied to quantify BRAF V600E and KRAS G12V in tumour-bearing nude mice serum, the results of which corresponded with those of real-time polymerase chain reaction. Overall, this study provides a promising alternative tool for the dynamic monitoring of ctDNA expression level which can benefit the clinical diagnosis of NSCLC. • CHA-induced powerful cycle amplification and the aggregation effect caused by MBs significantly enhanced the sensitivity of proposed pump-free SERS microfluidic chip, with the LODs low to aM level. • The pump-free SERS microfluidic chip presented satisfactory uniformity, stability, specificity and reproducibility. • This dual-signal amplification strategy could be applied for the dynamic monitoring expression of ctDNA in mice serum at different stages. [ABSTRACT FROM AUTHOR]