Precision Detection of Rifampin-resistant rpoB L378R Mutation in Mycobacterium tuberculosis with CRISPR-Cas12a

Abstract Rifampin is the most effective drug in the treatment of tuberculosis.However, certain strains of MTB have developed resistance to rifampin, leading to the need for alternative treatment options.The rpoB gene mutations play a central role in MTB resistance to the rifampin, so it is essential to identify these mutations and efficiently treat rifampin-resistant MTB strains.This study developed a novel CRISPR-Cas12a platform integrated with recombinase polymerase amplification (RPA) and fluorescence detection to specifically identify the rpoB_L378R mutation associated with Rifampin resistance in (MTB).We found that this detection system was highly specific and did not cross-react with created reference samples containing the genomes of MTB H37Rv, Mycobacterium smegmatis, Mycobacterium aureus, and Escherichia coli.The CRISPR-Cas12a-based platform developed in this study was simple, sensitive, and specific for detecting the Rifampin-resistant MTB strain with the rpoB_L378R mutation.This suggested that it has potential for clinical applications in identifying MTB rpoB_L378R mutation.