生物技术通报2025，Vol.41Issue(5)：62-69,8.DOI:10.13560/j.cnki.biotech.bull.1985.2024-0955
RPA-CRISPR/Cas12a结合重力驱动微流控芯片的MTB快检方法的建立
Gravity-driven Microfluidic Chip Based on RPA-CRISPR/Cas12a for the Rapid Detection of Mycobacterium tuberculosis
摘要
Abstract
[Objective]To develop a rapid and efficient method for detecting Mycobacterium tuberculosis(MTB),providing a technical solution for rapid screening of tuberculosis(TB)in grassroots units and remote areas,thereby enhancing TB prevention and control efforts.[Method]RPA(recombinase polymerase amplification)primers and crRNA were designed,and a gravity-driven microfluidic chip was introduced to establish an RPA-CRISPR/Cas12a on-chip detection method.The sensitivity and specificity of this method were further analyzed using samples from suspected TB patients for on-chip detection and sputum culture,comparing the established method with the sputum culture method for consistency.[Result]The established RPA-CRISPR/Cas12a on-chip detection method achieves MTB detection within 30 min,with a limit of detection(LOD)of 1 copy/μL.Using sputum culture identification as a reference standard,the on-chip detection method demonstrated a sensitivity of 91.11%,specificity of 94.34%,positive predictive value of 93.19%,negative predictive value of 92.59%,and accuracy of 92.86%,with a Kappa value of 0.856.[Conclusion]The developed RPA-CRISPR/Cas12a on-chip detection method presents high sensitivity and specificity,characterized by rapid,simple,and convenient.关键词
结核分枝杆菌/重组酶聚合酶扩增/CRISPR/Cas12a/微流控芯片/快速检测Key words
Mycobacterium tuberculosis/RPA/CRISPR/Cas12a/microfluidics chip/rapid detection引用本文复制引用
高畅,庄添驰,李宁,刘云,顾鹏飞,赵昕怡,季明辉..RPA-CRISPR/Cas12a结合重力驱动微流控芯片的MTB快检方法的建立[J].生物技术通报,2025,41(5):62-69,8.基金项目
江苏省卫生健康委员会预防医学科研课题面上项目(Ym2023064),国家级大学生创新创业训练计划基金项目(202410312005Z),南京预防医学科研课题(NJYFKT202409),江苏高校优势学科建设工程四期项目"护理学"(苏教研函[2023]11号) （Ym2023064）
