CAJ | 학술논문

环介导等温扩增技术(Loop-mediated isothermal amplification, LAMP)因其扩增速度快、灵敏度和特异性高、仪器要求低等优点而被广泛应用于核酸诊断领域。为充分利用 LAMP 技术优势、提高诊断检测的效率与可靠性、扩展其应用范围，同时节约试剂成本，近年来多重 LAMP技术的研究成为一大热点。常规的 LAMP扩增产物检测方法多数以聚合反应的双链DNA产物或其副产物为基础，只能判断有无扩增反应发生，而难以识别多重扩增产物的靶标来源及其特异性。为实现多重扩增产物的高特异检测，各国学者通过对该技术巧妙的改进或与其他技术相偶联，发展了一系列多重LAMP扩增检测技术。然而上述狭义的多重LAMP技术依然存在因引物间相互干扰、扩增效率存在差异而引发歧视性扩增的局限，限制了多重扩增的重数。近年研究活跃的微型扩增技术以其实现多个平行、互不干扰的小体积单重扩增的技术优势打破了这一局限，由此产生了新型的广义多重LAMP扩增技术。这些技术还具有试剂消耗少、自动化程度较高、交叉污染风险更小以及更适合对较多靶标进行现场快速检测等优势。本文分别从狭义多重 LAMP的方法原理及其扩增反应体系优化、广义多重 LAMP的方法原理以及多重LAMP技术在诊断检测中的应用等方面对近年来多重LAMP技术的研究进展进行了综述。
배개도등온확증기술(Loop-mediated isothermal amplification, LAMP)인기확증속도쾌、령민도화특이성고、의기요구저등우점이피엄범응용우핵산진단영역。위충분이용 LAMP 기술우세、제고진단검측적효솔여가고성、확전기응용범위，동시절약시제성본，근년래다중 LAMP기술적연구성위일대열점。상규적 LAMP확증산물검측방법다수이취합반응적쌍련DNA산물혹기부산물위기출，지능판단유무확증반응발생，이난이식별다중확증산물적파표래원급기특이성。위실현다중확증산물적고특이검측，각국학자통과대해기술교묘적개진혹여기타기술상우련，발전료일계렬다중LAMP확증검측기술。연이상술협의적다중LAMP기술의연존재인인물간상호간우、확증효솔존재차이이인발기시성확증적국한，한제료다중확증적중수。근년연구활약적미형확증기술이기실현다개평행、호불간우적소체적단중확증적기술우세타파료저일국한，유차산생료신형적엄의다중LAMP확증기술。저사기술환구유시제소모소、자동화정도교고、교차오염풍험경소이급경괄합대교다파표진행현장쾌속검측등우세。본문분별종협의다중 LAMP적방법원리급기확증반응체계우화、엄의다중 LAMP적방법원리이급다중LAMP기술재진단검측중적응용등방면대근년래다중LAMP기술적연구진전진행료종술。
Loop-mediated isothermal amplification (LAMP) has been widely applied in nucleic acid diagnostics due to its high sensitivity and specificity, high speed and low requirement of equipment. In order to fully leverage these merits, achieve high efficiency and reliability in diagnostics, and expand the applicable fields while keeping low reagent cost, multiplex LAMP technology has been extensively explored in recent years. Common methods for LAMP products detection are mostly based on the double-stranded DNA amplicons or byproducts from the polymerization reaction, so they can only identify the occurrence of amplification reaction but not the origins or specificity of the products. To achieve specific LAMP products detection, researchers developed various multiplex methods by im-proving the conventional LAMP technology or coupling LAMP with other assays. However, the interference and/or the different amplification efficiencies among different primer sets often lead to biased amplification and thus limited multiplexing level. We here defined these methods as narrow-sensed multiplex LAMP. The research on miniaturized amplification technology which is booming in recent years has given rise to the novel general-sensed multiplex LAMP technology that breaks this limitation by its capability to perform highly parallel and miniaturized simplex reactions in independent compartments. Methods of this type have additional benefits such as lower reagent cost, higher level of automation, lower risk of cross-contamination and better suitability for on-site detection of multiple targets. In this review, we summarize the recent research progress in multiplex LAMP technology from the following aspects: the principle and design of narrow-sensed LAMP and its amplification optimization, the general-sensed LAMP, and the various applications of all multiplex LAMP technologies in diagnostics.