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Dual-signal amplified self-powered biosensor integrating DNAzyme Walker and Dumbbell HCR for attomolar detection of TATA-28 DNA in thalassemia gene screening

  • 1Education Department of Guangxi Zhuang Autonomous Region, Laboratory of Optic-electric Chemo/Biosensing and Molecular Recognition, Engineering Research Center of Low-carbon and High-quality Utilization of Forest Biomass, University of Guangxi, Guangxi Key Laboratory of Chemistry and Engineering of Forest Products, Key Laboratory of Chemistry and Engineering of Forest Products, State Ethnic Affairs Commission, School of Chemistry and Chemical Engineering, Guangxi Minzu University, Nanning, 530006, China.
  • 2The Reproductive Hospital of Guangxi Zhuang Autonomous Region, Nanning, 530021, China.
  • 3Education Department of Guangxi Zhuang Autonomous Region, Laboratory of Optic-electric Chemo/Biosensing and Molecular Recognition, Engineering Research Center of Low-carbon and High-quality Utilization of Forest Biomass, University of Guangxi, Guangxi Key Laboratory of Chemistry and Engineering of Forest Products, Key Laboratory of Chemistry and Engineering of Forest Products, State Ethnic Affairs Commission, School of Chemistry and Chemical Engineering, Guangxi Minzu University, Nanning, 530006, China. Electronic address: gxunjww@126.com.
  • 4Education Department of Guangxi Zhuang Autonomous Region, Laboratory of Optic-electric Chemo/Biosensing and Molecular Recognition, Engineering Research Center of Low-carbon and High-quality Utilization of Forest Biomass, University of Guangxi, Guangxi Key Laboratory of Chemistry and Engineering of Forest Products, Key Laboratory of Chemistry and Engineering of Forest Products, State Ethnic Affairs Commission, School of Chemistry and Chemical Engineering, Guangxi Minzu University, Nanning, 530006, China. Electronic address: gxunxctan@126.com.
  • 5Education Department of Guangxi Zhuang Autonomous Region, Laboratory of Optic-electric Chemo/Biosensing and Molecular Recognition, Engineering Research Center of Low-carbon and High-quality Utilization of Forest Biomass, University of Guangxi, Guangxi Key Laboratory of Chemistry and Engineering of Forest Products, Key Laboratory of Chemistry and Engineering of Forest Products, State Ethnic Affairs Commission, School of Chemistry and Chemical Engineering, Guangxi Minzu University, Nanning, 530006, China. Electronic address: kejinghuang@163.com.
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June 14, 2025

Abstract

Thalassemia gene screening is vital for preventing severe infant morbidity, yet current methods lack the sensitivity for early clinical intervention. Here, we report an enzymatic biofuel cell self-powered biosensor (EBFC-SPB) integrating DNAzyme Walker and dumbbell hybridization chain reaction (DHCR) cascade amplification for attomolar-level detection of TATA-28, a critical thalassemia biomarker. The biosensor employs Au@Zr-MOF/graphdiyne (GDY) as a conductive substrate to immobilize glucose oxidase (bioanode) and [Ru(NH3)6]3+-responsive DNA circuits (biocathode). Target-activated DNAzyme Walker liberates single-stranded DNA (S0), initiating dumbbell HCR to generate electronegative DNA nanostructures that adsorb [Ru(NH3)6]3+. This process drives efficient electron transfer from the bioanode to the biocathode, greatly amplifying the open-circuit voltage (EOCV) compared to non-target conditions. The dual-amplification strategy achieves a linear response from 0.1 fM to 10 nM TATA-28 with a 35.3 aM detection limit (S/N = 3), surpassing existing methods in sensitivity. Successful validation in human serum (recovery: 90.1-106.5 %) highlights its clinical potential for early thalassemia screening.

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