A hairpin probe-mediated DNA circuit for the detection of the mecA gene of Staphylococcus aureus based on exonuclease III and DNAzyme-mediated signal amplification.

Abstract

A simple, highly sensitive biosensor for S. aureus detection is becoming increasingly important in human health and safety. In this work, a hairpin probe-mediated DNA circuit for the detection of the mecA gene of S. aureus was reported cascading Exo III-assisted cycling signal amplification and the DNAzyme-mediated cleavage reaction. In the presence of the target mecA gene, the recognition and hybridization between HP1 and mecA can trigger Exo III and DNAzyme-mediated signal amplification and further release numerous ATMND, resulting in an enhanced fluorescence response, which serves as a response signal for the fluorescence detection of mecA gene. This biosensor enables the sensitive and specific detection of the mecA gene, showing a linear response ranging from 1 fM to 1 nM with a detection limit of 0.5 fM. Moreover, this fluorescence assay has been applied for the analysis of clinical samples with satisfactory recovery. Importantly, this universal platform can be further extended for the analysis of other targets by alternating the corresponding recognition unit, which holds much promise in point-of-care testing for bacterial analysis.