Rapid Genotyping of the TPMT*3C Mutation Using Recombinase Polymerase Amplification and Lateral Flow Detection : A 20-Minute Isothermal Assay for Single Nucleotide Polymorphism (SNP) Genotyping of the TPMT*3C Mutation via Lateral Flow Readout for Thiopurine Dose Adjustment

Abstract

This study presents the development of a rapid genotyping assay for detecting the TPMT*3C mutation, a single nucleotide polymorphism (SNP) linked to increased risk of adverse reactions to thiopurine drugs. The assay combines recombinase polymerase amplification (RPA) with lateral flow detection, producing visually interpretable results in under 20 minutes using only basic heating equipment. Compared to conventional methods such as qPCR and LC-MS, this approach eliminates the need for complex instrumentation and trained personnel. Preliminary validation with nine clinical samples demonstrated 80% sensitivity and 100% specificity. These results highlight the feasibility of rapid, accessible TPMT*3C genotyping for pharmacogenomic dose adjustment, especially where laboratory infrastructure is limited.