Citation: QI Li-Juan, YANG Mei-Ting, DU Yan. Enzyme-Free Catalytic Hairpin Assembly Utilized in A Kinetically Competitive Aptasensor for Sensitive Detection of Ricin Toxin A Chain[J]. Chinese Journal of Analytical Chemistry, ;2021, 49(6): 1025-1034. doi: 10.19756/j.issn.0253-3820.211053 shu

Enzyme-Free Catalytic Hairpin Assembly Utilized in A Kinetically Competitive Aptasensor for Sensitive Detection of Ricin Toxin A Chain

QI Li-Juan ^1,2 ,
YANG Mei-Ting ^1,, ,
DU Yan ^1,2,,

1.
State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, China;
2.
School of Applied Chemistry and Engineering, University of Science & Technology of China, Hefei 230026, China

Corresponding author: YANG Mei-Ting, DU Yan,
Received Date: 21 January 2021
Revised Date: 12 April 2021

Fund Project: Supported by the National Natural Science Foundation of China (Nos.21874129, 31960506) and the International Scientific Cooperation Project of Jilin Scientific and Technological Development Program, China (No.20200801044GH).

A novel kinetically competitive aptasensor for detection of ricin toxin A chain (RTA) was fabricated based on the catalytic hairpin assembly (CHA) for signal amplification and transduction. The elements of kinetic competition contain aptamers (RA80), hairpin probes (HP1) and blockers, wherein the blockers kinetically compete with RA80 and HP1, but pre-binding of the target to its aptamer alters this kinetic competition and leads to more blocker binding to HP1. Then, more blockers binding to HP1 were separated as the catalyst for CHA reaction and increased fluorescence intensity. Thus, an indirect correlation between the fluorescence intensity and RTA concentration was constructed. Besides, the mismatched base pair introduced in the CHA circuit impeded uncatalyzed strand exchange reactions due to breathing or the mis-folding of nucleic acids into alternative conformers, which resulted in the decline of background signal and the improvement of signal-to-noise ratio. The limit of detection (LOD) for RTA was 0.88 nmol/L, which was comparable or even superior to other RTA determination strategies. The recoveries ranging from 91.2% to 107.9% were also satisfactory for RTA analysis in food powder. The proposed approach expanded the aptamer recognition mechanism and overcame the difficulty of the traditional antisense displacement strategy, bringing a bright hope to design and screen blockers for RNA aptamer with long sequence and complicated secondary structure without necessity of aptamer modification and requirement of lengthy pre-equilibration.
- Kinetic competition,
- Catalytic hairpin assembly,
- Aptasensor,
- Ricin toxin A chain,
- Detection
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