The Polymer Society of Korea

Abstract

Human erythropoietin (hEPO) enhances performance by increasing red blood cells and is banned by WADA due to its potential misuse for gene doping. DNA quantification is important for anti-doping test, but the conventional techniques rely on nucleic acid amplification which have inherent issues like and time-consuming and cross-contamination risk quantification bias. Here, we suggest a CRISPR-Cas12a based rapid detection technique for amplification-free and effective quantification of DNA at the amol level. To achieve this, we leveraged the fluid manipulation effect enabled by pneumatic valves to perform a Cas12a assay based on solution mixing within a loop reactor, which differs from conventional laminar-flow-based mixing strategies. This approach yielded excellent specificity and a rapid detection time of 30 minutes. Moreover, we demonstrated its versatility and quantification capability by directly detecting DNA from unpurified plasma samples using human pooled serum.