研究目的
To develop a surface-enhanced Raman scattering (SERS) aptasensor for the detection of saxitoxin, a harmful paralytic shellfish toxin, using an aptamer as the recognition element and crystal violet as a label-free Raman reporter, aiming for high sensitivity, selectivity, and simplicity in on-site applications.
研究成果
The developed SERS aptasensor successfully detects saxitoxin with high selectivity and a linear range of 10-200 nM, demonstrating good accuracy and reproducibility in real shellfish samples. It offers a simple, label-free approach but requires optimization for temperature stability and could be improved for lower detection limits. Future work should focus on enhancing sensitivity and adapting the method for broader environmental monitoring.
研究不足
The SERS aptasensor requires detection at high temperatures (68°C), which may complicate on-site use. It has a limit of detection of 11.7 nM, which is not as low as some other methods (e.g., ELISA). The method may be affected by matrix components in real samples, though background subtraction was used. Larger gold nanoparticles (e.g., 49 nm) are unstable and precipitate quickly.
1:Experimental Design and Method Selection:
The study designed an SERS aptasensor using gold nanoparticles functionalized with a saxitoxin-specific aptamer (M-30f) and crystal violet as a Raman reporter. The method leverages the conformational changes in the aptamer induced by saxitoxin at high temperatures to modulate SERS signals.
2:Sample Selection and Data Sources:
Saxitoxin standards and real shellfish samples were used. Shellfish were homogenized, spiked with saxitoxin, and extracted using acetic acid, following AOAC procedures.
3:List of Experimental Equipment and Materials:
Gold nanoparticles synthesized from chloroauric acid and trisodium citrate, oligonucleotides (M-30f, SH-M-30f, T5), crystal violet, saxitoxin and analogs (neosaxitoxin, gonyautoxin 2&3), buffers (phosphate, Tris-HCl, acetate, tris-acetate), and various reagents. Equipment included a transmission electron microscope (JEOL-2100F), ultraviolet-visible spectrophotometer (Agilent Cary 300), Raman spectrometer (Horiba Jobin Yvon LabRAM HR Evolution), and centrifugation systems.
4:Experimental Procedures and Operational Workflow:
Gold nanoparticles were synthesized and characterized. They were functionalized with DNA via gold-sulfur bonds. SERS assays involved mixing gold nanoparticle-DNA conjugates with crystal violet and saxitoxin, incubating, adding NaCl to induce aggregation, and measuring Raman spectra at optimized temperatures. Real sample analysis involved extraction and similar SERS measurements.
5:Data Analysis Methods:
Raman spectra were analyzed using LabSpec 6 software for background subtraction. Intensity at 1187 cm?1 was used for quantification. Statistical analysis included linear regression for sensitivity and standard deviation for reproducibility.
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