DNAzyme Sensor for the Detection of Ca²⁺ Using Resistive Pulse Sensing

• Main Authors: Imogen Heaton, Mark Platt
• Format: Article
• Language: English
• Published: MDPI AG 2020-10-01
• Series: Sensors
• Subjects: DNAzyme; aptamer; nanopore; resistive pulse sensor; metal ion sensor
• Online Access: https://www.mdpi.com/1424-8220/20/20/5877
• ISSN: 1424-8220

DNAzymes are DNA oligonucleotides that can undergo a specific chemical reaction in the presence of a cofactor. Ribonucleases are a specific form of DNAzymes where a tertiary structure undergoes cleavage at a single ribonuclease site. The cleavage is highly specificity to co-factors, which makes them excellent sensor recognition elements. Monitoring the change in structure upon cleavage has given rise to many sensing strategies; here we present a simple and rapid method of following the reaction using resistive pulse sensors, RPS. To demonstrate this methodology, we present a sensor for Ca²⁺ ions in solution. A nanoparticle was functionalised with a Ca²⁺ DNAzyme, and it was possible to follow the cleavage and rearrangement of the DNA as the particles translocate the RPS. The binding of Ca²⁺ caused a conformation change in the DNAzyme, which was monitored as a change in translocation speed. A 30 min assay produced a linear response for Ca²⁺ between 1–9 μm, and extending the incubation time to 60 min allowed for a concentration as low as 0.3 μm. We demonstrate that the signal is specific to Ca²⁺ in the presence of other metal ions, and we can quantify Ca²⁺ in tap and pond water samples.