Coupling Exponential to Linear Amplification for Endpoint Quantitative Analysis

Abstract Exponential DNA amplification techniques are fundamental in ultrasensitive molecular diagnostics. These systems offer a wide dynamic range, but the quantification requires real‐time monitoring of the amplification reaction. Linear amplification schemes, despite their limited sensitivity, can achieve quantitative measurement from a single end‐point readout, suitable for low‐cost, point‐of‐care, or massive testing. Reconciling the sensitivity of exponential amplification with the simplicity of end‐point readout would thus break through a major design dilemma and open a route to a new generation of massively scalable quantitative bioassays. Here a hybrid nucleic acid‐based circuit design is introduced to compute a logarithmic function, therefore providing a wide dynamic range based on a single end‐point measurement. CELIA (Coupling Exponential amplification reaction to LInear Amplification) exploits a versatile biochemical circuit architecture to couple a tunable linear amplification stage – optionally embedding an inverter function – downstream of an exponential module in a one‐pot format. Applied to the detection of microRNAs, CELIA provides a limit of detection in the femtomolar range and a dynamic range of six decades. This isothermal approach bypasses thermocyclers without compromising sensitivity, thereby opening the way to applications in various diagnostic assays, and providing a simplified, cost‐efficient, and high throughput solution for quantitative nucleic acid analysis.