Work

Universal all-in-one hyper-branched rolling circle amplification based biosensor for quantitative and qualitative detection of nucleic acids

Year: 2025

Type: article

Abstract: Universal, simple, sensitive, and specific approaches for the quantitative and qualitative detection of nucleic acids are urgently needed in clinical diagnosis and biological detection. Therefore, in this study, an universal all-in-one hyper-branched rolling circle amplification (HRCA) based biosensor (referred to as the all-in-one HRCA biosensor) was developed for highly sensitive and specific nucleic acid quantification and quantitative detection using an elaborately screened detection set, which includes a padlock probe, forward primer, reverse primer, and molecular beacon (MB). The maximal similarity of the detection set to the human genome and transcriptome is less than 75%, and there is no existing secondary structure, ensuring high specificity and sensitivity of the all-in-one HRCA biosensor. The developed all-in-one biosensor exhibited a broad linear dynamic range of 1 fM to 0.1 nM and high specificity, enabling discrimination of single base differences in the target sequence. The test results of real samples showed that the established all-in-one HRCA biosensor could be used for qualitative or quantitative detection of nucleic acids in cells, throat swabs, and plasma samples, demonstrating high accuracy. Therefore, we envision that the all-in-one HRCA biosensor can serve as a robust assay for universal, sensitive and specific detection of various nucleic acid targets, as well as clinical molecular diagnosis.

Source: Sensing and Bio-Sensing Research

DOI: 10.1016/j.sbsr.2025.100829