ABSTRACT
A dual-mode nanoprobe was constructed to detect Bax messenger RNA (mRNA), consisting of gold nanotriangles (AuNTs), a Cy5-modified recognition sequence, and a thiol-modified DNA sequence. Bax mRNA is one of the key pro-apoptotic factors in the apoptosis pathway. Raman enhancement and fluorescence quenching of the signal group Cy5 were performed using AuNTs as substrates. The thiol-modified nucleic acid chain is partially complementary to the Cy5-modified nucleic acid chain to form a double strand and is linked to the AuNTs by the Au-S bond. When Bax mRNA is present, the Cy5-modified strand specifically binds to it to form a more stable duplex, making Cy5 far away from AuNTs, and SERS signal is weakened while fluorescence signal is enhanced. The nanoprobe can be used for the quantitative detection of Bax mRNA in vitro. Combined with the high sensitivity of SERS and the visualization of fluorescence, this method has good specificity and can be used for in situ imaging and dynamic monitoring of Bax mRNA during deoxynivalenol (DON) toxin-induced apoptosis of HepG2 cells. DON plays a pathogenic role mainly by inducing cell apoptosis. The results confirmed that the proposed dual-mode nanoprobe has good versatility in various human cell lines.
Subject(s)
Apoptosis , Sulfhydryl Compounds , Humans , bcl-2-Associated X Protein , RNA, Messenger , Fluorescence , Cell Line, TumorABSTRACT
Histamine is a significant biomarker to assess the freshness of fish products. In this study, a novel MOF-based SERS sensor for histamine determination was synthesized by wrapping PVP-capped Au nanoflowers with a ZIF-67 shell (Au NFs@ZIF-67). The highly branched Au NFs core exhibited a strong electromagnetic field enhancement effect and provided an ultra-sensitive SERS fingerprint spectrum, while ZIF-67 shell was the contributor to enrich the target and stabilize the substrate. The morphology of the core-shell structures can be easily controlled by the concentrations of the capping agent PVP and MOF precursor Co ion. Consequently, 4-MBA pre-grafted on the optimized SERS substrate can act as the Raman internal standard (IS) to eliminate signal fluctuations through standardizing all spectra against its peak at 1074 cm-1. Moreover, as the specific receptor for histamine molecules, 4-MBA helped reach the low detection sensitivity, where the SERS intensity ratio, I1172/I1074 presented a good linear relationship towards the histamine concentrations (10-3-10-7 M) with the LOD of 0.87 × 10-7 M (R2 = 0.9930). Furthermore, the application in monitoring fish spoilage process demonstrated the feasibility and reliability of the developed sensor. This work provided a facile strategy to construct MOF-based SERS substrate as a potential platform for the shelf-life prediction of fish products.
Subject(s)
Metal Nanoparticles , Nanocomposites , Animals , Histamine , Reproducibility of Results , Sulfhydryl Compounds/chemistry , Fishes , Spectrum Analysis, Raman , Gold/chemistry , Metal Nanoparticles/chemistryABSTRACT
Salmonella typhimurium (S. typhimurium) and Staphylococcus aureus (S. aureus) are the two most important foodborne pathogens which can easily cause disease infections. Here, the aptamer-facilitated gold/silver nanodimer SERS probes were built for the simultaneous detection of the two bacteria with the help of magnetic separation enrichment. First, two nanodimer SERS signal probes and two magnetic capture probes each connected with the specific aptamer were fabricated. The distance between gold and silver nanoparticles in the dimer can amplify the Raman signal (Cy3 and Rox) at the junction but modified in the aptamer sequence. Then, after the addition of S. typhimurium and S. aureus, the sandwich-like composite structures "SERS signal probes-target-magnetic capture probes" formed because of the high affinity between aptamer sequences and their target bacteria. Under the optimal experimental conditions, the linear correlations between Raman intensity and the logarithm of the concentration of bacteria were y = 876.95x-67.84 (R2 = 0.9865) for S. typhimurium and y = 1280.43x-1752.6 (R2 = 0.9883) for S. aureus. The SERS detection showed the nanodimer probe had high selectivity. Besides, the recovery experiment in milk sample indicated good accuracy compared with the traditional plate counting method.