研究目的
To develop a rapid method for identification and typing of lipopolysaccharides (LPS) using dual aptamer-functionalized gold nanoparticles for colorimetric detection.
研究成果
The developed colorimetric method using dual aptamer-functionalized AuNPs allows for rapid, visible detection and typing of LPS with high specificity and a detection limit of 1 μg·mL?1. It offers advantages such as no need for enzymes or toxic reagents, cost-effectiveness, and potential applications in medical diagnostics, environmental monitoring, and food safety.
研究不足
The method is specific to the aptamers used and may not detect other types of LPS or bacteria without further aptamer development. The detection limit is 1 μg·mL?1, which might not be sensitive enough for very low concentrations. Optimization of binding buffers was required, indicating sensitivity to experimental conditions.
1：Experimental Design and Method Selection:
Utilized gold nanoparticles (AuNPs) functionalized with aptamers for colorimetric detection based on aggregation-induced color change. Designed a logic typing method using a general probe (G-probe) for ethanolamine and a specific probe (S-probe) for E. coli O111:B4 LPS.
2：Sample Selection and Data Sources:
Used E. coli O111:B4 LPS and E. coli O55:B5 LPS as model analytes. DNA oligonucleotides were ordered from Invitrogen Trading Co., Ltd.
3：List of Experimental Equipment and Materials:
Equipment includes Thermo Scientific Varioskan Flash for colorimetric measurements, digital camera (MEIZU MX), JOEL Transmission Electron Microscope JEM2100. Materials include HAuCl4, Tris-base, TCEP, LPS samples, and aptamers.
4：Materials include HAuCl4, Tris-base, TCEP, LPS samples, and aptamers. Experimental Procedures and Operational Workflow:
4. Experimental Procedures and Operational Workflow: Functionalized AuNPs with aptamers, optimized binding buffer conditions, added LPS samples to probes, monitored color changes and absorbance at 524 nm, and performed TEM imaging.
5：Data Analysis Methods:
Measured optical density changes at 524 nm, generated calibration curves for quantitative analysis, and used visual inspection for typing.
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Varioskan Flash
Thermo Scientific Varioskan Flash
Thermo Fisher Scientific
Colorimetric measurements
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Transmission Electron Microscope
JEM2100
JOEL
Imaging to confirm aggregation state of AuNPs
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Chloroauric Acid
HAuCl4
Sigma-Aldrich
Precursor for synthesizing gold nanoparticles
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Tris (2-carboxyethyl) phosphine
TCEP
Sigma-Aldrich
Reducing agent used in aptamer functionalization
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E. coli LPS
O111:B4 and O55:B5
Sigma-Aldrich
Target analytes for detection and typing
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Digital Camera
MEIZU MX
MEIZU
Photo collection for visual inspection of color changes
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Gold Nanoparticles
Core component for colorimetric sensing, functionalized with aptamers
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DNA Oligonucleotides
Invitrogen
Aptamers used for functionalizing AuNPs
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Tris-base
Sigma-Aldrich
Component of binding buffers
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