A Multitargeted Electrochemiluminescent Biosensor Coupling DNAzyme with Cascading Amplification for Analyzing Myocardial miRNAs

Abstract

Several circulating miRNAs are associated with the pathogenic process of acute myocardial infarction (AMI). Thus, analyzing myocardial miRNAs in the circulatory system is important for the diagnosis and treatment of AMI, especially for early-stage diagnosis. Based on the characteristics of myocardial miRNAs, an ultrasensitive and multitargeted electrochemiluminescence (ECL) sensing platform was developed with a versatile probe that can couple DNAzyme with hybridization chain reaction amplification. The target miRNA and auxiliary chains form a circular unit that shears the versatile probe hairpin, and the products subsequently trigger cascading amplification; a long strand of dsDNA is then generated with many C-rich sequences that can undergo in situ reductions to generate ECL luminophore silver clusters. Using this strategy, three myocardial miRNAs are successfully detected with a detection limit as low as 29.6 aM (S/N = 3). Notably, our method can detect myocardial miRNA groups composed of multiple related circulating miRNAs with high selectivity over interfering miRNAs in blood. This is extremely important for solving the problem of diverse and low abundance of infarct-associated miRNAs. Our strategy pioneers a new idea of miRNA detection, and given its versatility and sensitivity, it is promising for the diagnosis of multigene-regulated cardiovascular diseases.

Anti-Fouling Magnetic Beads Combined with Signal Amplification Strategies for Ultra-Sensitive and Selective Electrochemiluminescence Detection of MicroRNAs in Complex Biological Media

“…Classical PCR amplification and microarrays are traditional miRNA analysis strategies. − In recent years, analysis strategies for miRNA research such as surface-enhanced Raman, miRNA imaging, electrochemiluminescence (ECL) sensors, and electrochemical sensors have also been reported. − However, due to the similarity of homologous miRNAs and the problems of trace amounts in biological samples, these methods inevitably suffer from miRNA detection errors, low sensitivity, or complicated operations. Therefore, the introduction of signal amplification design in the detection system has become a crucial step.…”

Multistage nucleic acid amplification induced nano-aggregation for 3D hotspots-improved SERS detection of circulating miRNAs

“…Relevant nucleic acid structures were designed based on our previous work [19,33,38,39]. Characterization of target recognition and the first amplification step with DNAzyme were performed using polyacrylamide gel electrophoresis (10% w/w gel) after incubating the samples at 37 °C for 9 h (10 mM MgAC 2 , TE buffer).…”

“…The focus of our studies is on miRNA-499, miRNA-328, and miRNA-208, because the levels of these miRNAs are robustly increased in the blood circulation after myocardial infarction [31,32]. Accurate identification and efficient capture of miRNAs were achieved using an assembled DNAzyme structure that we have reported previously [33]. The principle of the SERS platform in detecting circulating miRNA is shown in Scheme 1.…”

Multistage Nucleic Acid Amplification Induced Nano-Aggregation for 3D Hotspots-Improved SERS Detection of Circulating miRNAs

“…Relevant nucleic acid structures were designed based on our previous work [19,33,38,39]. Characterization of target recognition and the rst ampli cation step with DNAzyme were performed using polyacrylamide gel electrophoresis (10% w/w gel) after incubating the samples at 37•C for 9 h (10 mM MgAC2, TE buffer).…”

“…The focus of our studies is on miRNA-499, miRNA-328, and miRNA-208, because the levels of these miRNAs are robustly increased in the blood circulation after myocardial infarction [31,32]. Accurate identi cation and e cient capture of miRNAs were achieved using an assembled DNAzyme structure that we have reported previously [33]. The principle of the SERS platform in detecting circulating miRNA is shown in Scheme 1.…”