A novel chemiluminescent peroxidase-like DNA nanomachine for the detection of bacterial nucleic acids

Accurate and rapid nucleic acid detection is crucial for safeguarding public health across diverse applications, including healthcare, environmental monitoring, and food safety. This study introduces a novel chemiluminescent peroxidase-like DNA nanomachine (PxDm) system, integrated within a compact, closed-type device, enabling a complete workflow from sample introduction to result readout. This system was designed for the detection of bacterial nucleic acids, specifically targeting the 16S rRNA of E.coli. Notably, the PxDm system demonstrates versatility, effectively detecting target sequences across varying sample purities, including synthetic single- and double-stranded DNA, PCR and DAMP amplicons, extracted RNA, and crude cell lysates. The PxDm for E. coli exhibited a limit of detection of 12 nM with a selectivity factor 86% against non-target microorganisms. This compact and deliverable device facilitates rapid detection, achieving a limit of detection of 155 nM for double-stranded sequences. The PxDm system's ability to function across a range of sample types without extensive purification underscores its potential for diverse applications, including rapid diagnostics and food safety monitoring.