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An aptamer-based single particle method for sensitive detection of thrombin using fluorescent quantum dots as
Jinjin Yin1, Aidi Zhang1, Chaoqing Dong1
1School of Chemistry and Chemical Engineering, State Key Laboratory of Metal Matrix Composites, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai 200240, PR China.
Talanta
|October 11, 2015
Summary
This study presents a new aptamer-based method using quantum dots and fluorescence correlation spectroscopy for detecting thrombin in human serum. The assay demonstrates high sensitivity and specificity for thrombin detection.
Area of Science:
- Biomedical Engineering
- Analytical Chemistry
- Biophysics
Background:
- Thrombin is a key biomarker in various physiological and pathological processes.
- Accurate and sensitive detection of thrombin in biological samples is crucial for clinical diagnostics.
- Existing detection methods may lack sensitivity, specificity, or require complex sample preparation.
Purpose of the Study:
- To develop a novel aptamer-based single particle method for sensitive thrombin detection.
- To utilize fluorescence correlation spectroscopy (FCS) combined with quantum dots (QDs) for real-time thrombin quantification.
- To validate the method's performance in human serum samples.
Main Methods:
- Development of a single particle assay using quantum dots (QDs) labeled with thrombin-binding aptamer (TBA).
- Employing fluorescence correlation spectroscopy (FCS) to monitor changes in QD diffusion time upon thrombin binding.
- Optimization of assay conditions to enhance sensitivity and specificity.
- Validation using total internal reflection fluorescence microscopy (TIRFM) imaging.
Main Results:
- Thrombin binding induced QD dimerization, leading to a measurable increase in diffusion time detected by FCS.
- Optimized assay achieved a linear detection range from 5.0 nM to 500 nM for thrombin.
- The limit of detection (LOD) for thrombin was determined to be approximately 2.6 nM.
- Successful application of the method for homogeneous thrombin determination in human serum samples.
Conclusions:
- The developed aptamer-based single particle method provides a sensitive and specific approach for thrombin detection.
- Fluorescence correlation spectroscopy combined with QDs offers a powerful tool for analyzing molecular interactions in complex biological fluids.
- This method holds potential for clinical diagnostics and research applications requiring accurate thrombin quantification.
