Highly sensitive biosensor for specific miRNA detection based on cascade signal amplification and magnetic...

Herein, magnetic electrochemiluminescence (ECL) nanoparticle Fe₃O₄@PtPd/Ru(bpy)₃²⁺ had been synthesized then been coupled with CRISPR/Cas13a system and Zn²⁺ dependent DNAzyme to design a novel ECL biosensor for specific detection of microRNA-145 (miRNA). The synthesized multifunctional magnetic nanoluminescent materials Fe₃O₄@PtPd/Ru(bpy)₃²⁺ not only load Ru(bpy)₃²⁺ to provide ECL signals, but also can quickly achieve separation and enrichment from complex matrices. In addition, ferrocene (Fc) was used as a quencher in the Ru(bpy)₃²⁺/tripropylamine (TPA) system. Fc was modified on DNA bound to Fe₃O₄@PtPd. Benefited from the highly specific recognition ability of CRISPR/Cas13a, the target miRNA induces CRISPR/Cas13a trans-cleavage to trigger the Zn²⁺-dependent DNAzyme cyclic cleavage to realize the dual signal amplification. DNA modified by Fc was split by target miRNA-induced cleaving, and then magnetic separation was performed to keep Fc away from the surface of the nanoparticles. Thus, the enhanced ECL signal was obtained to detect miRNA-145. Under optimized conditions, the prepared sensor showed a wide linear range (1 fM to 1 nM) and a low limit of detection (LOD) down to 0.41 fM. Furthermore, it shows excellent selectivity and good reproducibility. The proposed ECL platform has huge potential applications in the development of various sensitive sensors for detecting the other miRNA.