研究目的
To develop a sensitive and accurate method for the detection of 17β-estradiol using surface-enhanced Raman spectroscopy (SERS) combined with hybridization chain reaction (HCR) amplification on Au@Ag core-shell nanoparticles, addressing the need for trace analysis in environmental and food safety contexts.
研究成果
The developed aptamer-based SERS method with HCR amplification provides high sensitivity, specificity, and reproducibility for 17β-estradiol detection, with a low detection limit of 0.1 pM and successful application in real urine samples. It shows promise for trace analysis but requires simplification for practical use.
研究不足
The two-step signal amplification process is described as cumbersome, and further optimization into a simpler, one-step or one-pot model is needed. The method may require refinement for broader application in complex systems.
1：Experimental Design and Method Selection:
The assay combines SERS with HCR for signal amplification. Au@Ag core-shell nanoparticles are used as SERS substrates due to their enhanced plasmonic properties. HCR is employed as an enzyme-free amplification strategy to assemble multiple nanoparticles and amplify the Raman signal.
2：Sample Selection and Data Sources:
Synthetic oligonucleotides (aptamers and probes) are used, and real samples include spiked human urine. Samples are prepared by dissolving 17β-estradiol in acetone and diluting with urine to various concentrations.
3：List of Experimental Equipment and Materials:
Key materials include HAuCl4·3H2O, sodium citrate, AgNO3, R6G, oligonucleotides, nicking enzyme N.BstNBI, streptavidin, and microtiter plates. Equipment includes a LabRAM HR Evolution Raman spectrometer for measurements.
4：Experimental Procedures and Operational Workflow:
Steps involve synthesis of Au@Ag nanoparticles, labeling with R6G, conjugation with DNA probes, HCR initiation in microplate wells, and Raman measurement. Optimization of probe ratios and conditions is performed.
5：Data Analysis Methods:
Raman spectra are analyzed, with intensity at 1651 cm?1 used for quantification. Statistical analysis includes calculation of detection limits, linear range, and recovery rates from spiked samples.
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LabRAM HR Evolution
HR Evolution
HORRIBA JOBIN YVON
Raman spectrometer used for measuring SERS signals in the experiment.
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Gold (III) chloride trihydrate
HAuCl4·3H2O
Sangon
Chemical used for synthesizing gold nanoparticles.
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Sodium citrate dehydrate
Na3C6H5O7·2H2O
Sangon
Reducing agent used in nanoparticle synthesis.
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Silver nitrate
AgNO3
Sangon
Used for forming silver shell on gold nanoparticles.
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Rhodamine 6G
R6G
Sangon
Raman reporter molecule adsorbed on nanoparticles for SERS detection.
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Nicking enzyme N.BstNBI
N.BstNBI
Sangon
Endonuclease used to hydrolyze specific DNA sequences in the assay.
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Streptavidin
Sangon
Used for immobilizing biotinylated probes on microplate wells.
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Microtiter plate
96-well
Platform for conducting the hybridization chain reaction and Raman measurements.
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