Yang Nan, Zhang Han, Han Xiu, Liu Zhifeng, Lu Yan
Department of Gastroenterology, Children's Hospital of Nanjing Medical University, Nanjing, China.
Institute of Translational Medicine, Medical College, Yangzhou University, Yangzhou, China.
Front Microbiol. 2024 Jul 17;15:1406632. doi: 10.3389/fmicb.2024.1406632. eCollection 2024.
Loop-mediated isothermal amplification (LAMP) is a novel method for nucleic acid detection known for its isothermal properties, high efficiency, sensitivity, and specificity. LAMP employs 4 to 6 primers targeting 6 to 8 regions of the desired sequence, allowing for amplification at temperatures between 60 and 65°C and the production of up to 10 copies within a single hour. The product can be monitored by various methods such as turbidimetry, fluorometry, and colorimetry. However, it faces limitations such as the risk of non-specific amplification, challenges in primer design, unsuitability for short gene sequences, and difficulty in multiplexing. Recent advancements in polymerase and primer design have enhanced the speed and convenience of the LAMP reaction. Additionally, integrating LAMP with technologies like rolling circle amplification (RCA), recombinase polymerase amplification (RPA), and CRISPR-Cas systems has enhanced its efficiency. The combination of LAMP with various biosensors has enabled real-time analysis, broadening its application in point-of-care testing (POCT). Microfluidic technology has further facilitated the automation and miniaturization of LAMP assays, allowing for the simultaneous detection of multiple targets and preventing contamination. This review highlights advancements in LAMP, focusing on primer design, polymerase engineering, and its integration with other technologies. Continuous improvements and integration of LAMP with complementary technologies have significantly enhanced its diagnostic capabilities, making it a robust tool for rapid, sensitive, and specific nucleic acid detection with promising implications for healthcare, agriculture, and environmental monitoring.
环介导等温扩增技术（LAMP）是一种新型核酸检测方法，以其等温特性、高效性、灵敏度和特异性而闻名。LAMP使用4至6条引物靶向目标序列的6至8个区域，可在60至65°C的温度下进行扩增，1小时内可产生多达10个拷贝。产物可通过多种方法进行监测，如比浊法、荧光法和比色法。然而，它也面临一些局限性，如非特异性扩增风险、引物设计挑战、不适用于短基因序列以及多重检测困难等。聚合酶和引物设计方面的最新进展提高了LAMP反应的速度和便利性。此外，将LAMP与滚环扩增（RCA）、重组酶聚合酶扩增（RPA）和CRISPR-Cas系统等技术相结合提高了其效率。LAMP与各种生物传感器的结合实现了实时分析，拓宽了其在即时检测（POCT）中的应用。微流控技术进一步促进了LAMP检测的自动化和小型化，可同时检测多个目标并防止污染。本综述重点介绍了LAMP的进展，包括引物设计、聚合酶工程及其与其他技术的整合。LAMP与互补技术的持续改进和整合显著增强了其诊断能力，使其成为一种用于快速、灵敏和特异性核酸检测的强大工具，对医疗保健、农业和环境监测具有重要意义。
