Journal
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
Volume 193, Issue 10, Pages 3139-3150Publisher
SPRINGER
DOI: 10.1007/s12010-021-03585-x
Keywords
Beclomethasone; Systematic evolution of ligands by exponential enrichment; SELEX; Aptamers; Quantum dot-based assay
Funding
- Guangdong Biomedical Technology Collaborative Innovation Center [ZA20190010]
- Demonstration Base of Achievement Transformation of Rapid Detection Methods for Health Products [ZA20180049]
- Key Laboratory of Quick Screening and Quick Inspection of Health Products [ZA20180046]
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The research team successfully developed an aptamer specific for beclomethasone (BEC) and a quantum dot-based assay (QDA) for BEC detection. The QDA method showed high sensitivity and accuracy, suitable for detecting BEC in Traditional Chinese Medicine (TCM).
We developed an aptamer that was specific for beclomethasone (BEC) via systematic evolution of ligands by exponential enrichment (SELEX). Development was monitored by real-time quantitative PCR (Q-PCR) and the enriched library was sequenced by high-throughput sequencing. Forty-seven aptamer candidates were obtained; of these, BEC-6 showed the highest affinity (K-d = 0.15 +/- 0.02 mu M) and did not cross-react with other BEC analogs. We also developed a quantum dot-based assay (QDA) for the detection of BEC that was based upon a quantum dot (QD) composite probe. Under optimized reaction conditions, the linear range of this method for BEC was 0.1 to 10 mu M with a low detection limit (LOD) of 0.1 mu M. Subsequently, the method was used to detect BEC in Traditional Chinese Medicine (TCM) with a mean recovery of 81.72-91.84%. This is the first report to describe the development of an aptamer against BEC; BEC-6 can also be engineered into QDA for the detection of BEC.
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