研究目的
To construct a new class of DNA logic gates for simultaneous detection of Hg2+ and Pb2+ ions using a SERS-active platform based on DNAzyme activity dependent on metal ions, aiming for high sensitivity and specificity in environmental and analytical applications.
研究成果
The SERS-active platform successfully enables sensitive and specific simultaneous detection of Hg2+ and Pb2+ ions with low detection limits, and it introduces a novel DNA logic gate concept. This approach is promising for environmental monitoring and can be extended to other metal ions and applications.
研究不足
The method may have limitations in complex matrices due to potential interferences, and the DNAzyme activity might be affected by environmental factors. The detection limits, while low, could be further optimized, and the system's applicability to other metal ions requires additional validation.
1：Experimental Design and Method Selection:
The study employs a SERS-based biosensor using oligonucleotide-functionalized gold-coated polystyrene microspheres. DNAzyme activity is leveraged for metal ion detection, with specific DNA sequences designed for Hg2+ and Pb2+ recognition.
2：Sample Selection and Data Sources:
Synthetic DNA strands (S1, S2, S3, S4) are used, along with standard solutions of Hg2+ and Pb2+ ions. Real water samples (lake water and tap water) are spiked with known concentrations for validation.
3：List of Experimental Equipment and Materials:
Polystyrene microspheres, gold nanoparticles, DNA strands, Raman reporters (DTNB and 4-MBA), chemicals like HAuCl4, trisodium citrate, PBS buffer, BSA, TCEP, and various metal ions for specificity tests. Equipment includes SEM, TEM, Raman microscope (in Via, Renishaw), vapor deposition system, centrifuge, and water purification system (ELGA).
4：Experimental Procedures and Operational Workflow:
Preparation of polystyrene microspheres and gold-coated substrates, functionalization with DNA, fabrication of Au nanoconjugates, SERS detection by mixing samples with functionalized nanoparticles, incubation, washing, and Raman spectroscopy measurement.
5：Data Analysis Methods:
SERS signal intensity is measured at specific Raman shifts (1330 cm-1 for Hg2+, 1580 cm-1 for Pb2+), with calibration curves constructed for quantitative analysis using linear regression. Specificity is tested against other metal ions, and recovery studies are conducted in real samples.
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