Chain extension-mediated colorimetric and sensitive detection of osteosarcoma-related miRNA based on CHA-cooperated self-priming

Abstract

Abstract The accurate and dynamic monitoring of osteosarcoma, one of the most prevalent cancers, has a substantial impact on the quality of life of numerous individuals. This study proposes a sample, sensitive, and portable approach to miRNA detection that integrates a catalytic hairpin assembly (CHA) circuit with self-priming mediated chain extension. This method employs CHA as a protein enzyme-free isothermal amplification technique to amplify miRNA signals and induce self-priming through a specially designed hairpin probe. The pyrophosphate sensing probe (pp probe) was employed, which enables dual high-efficiency and stable colorimetric signaling, in contrast to traditional colorimetric modes. This novel system for the colorimetric detection of target miRNA is operated with high sensitivity (LOD of 3.5 fM) and selectivity, with the entire detection procedure requiring approximately 60 min. Additionally, this system could detect miRNA from constructed serum samples with high stability and recovery. This system is unique, extremely simple, and rapid, and it is designed to detect osteosarcoma-related miRNA through a highly sensitive color change. It is likely to be useful in applications that necessitate point-of-care detection of osteosarcoma.

Keywords

• microRNA
• Catalytic hairpin assembly
• Colorimetric assay
• DNA polymerase