多重环介导等温扩增技术研究进展

doi:10.16288/j.yczz.15-141

遗传 ›› 2015, Vol. 37 ›› Issue (9): 899-910.doi: 10.16288/j.yczz.15-141

多重环介导等温扩增技术研究进展

林文慧^{1, 2}, 邹秉杰², 宋沁馨¹, 周国华^{1, 2, 3}

1.中国药科大学,生命科学与技术学院,南京210009;
2.南京大学医学院附属金陵医院,药理科,南京210002;
3.南京大学生命分析化学国家重点实验室,南京210008

收稿日期:2015-04-03 修回日期:2015-05-10 出版日期:2015-09-20 发布日期:2015-09-20
通讯作者: 周国华,博士,研究员,研究方向：分子诊断。E-mail: ghzhou@nju.edu.cn
作者简介:林文慧,硕士研究生,专业方向：分子诊断。E-mail: fjaxlwh@163.com
基金资助:
国家自然科学基金项目(编号：31200638, 31300704，21275161),江苏省临床科技专项项目(编号：BL2012061)和国家重点科学技术专项项目(编号：2013ZX10004103-001)资助

Progress in multiplex loop-mediated isothermal amplification technology

Wenhui Lin^{1, 2}, Bingjie Zou², Qinxin Song¹, Guohua Zhou^{1, 2, 3}

1. School of Life Science and Technology, China Pharmaceutical University, Nanjing 210009,China;
2.Department of Pharmacology, Jinling Hospital, Medical School of Nanjing University, Nanjing 210002,China;
3.State Key Laboratory of Analytical Chemistry for Life Science, Nanjing University, Nanjing 210008,China

Received:2015-04-03 Revised:2015-05-10 Online:2015-09-20 Published:2015-09-20

摘要/Abstract

摘要： 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。

关键词: 多重环介导等温扩增, 特异扩增产物检测, 微型扩增技术, 微流控芯片

Abstract: 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 improving 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.

Key words: multiplex loop-mediated isothermal amplification, specific amplification product detection, miniaturized amplification technology, microfluidic chip

林文慧, 邹秉杰, 宋沁馨, 周国华. 多重环介导等温扩增技术研究进展[J]. 遗传, 2015, 37(9): 899-910.

Wenhui Lin, Bingjie Zou, Qinxin Song, Guohua Zhou. Progress in multiplex loop-mediated isothermal amplification technology[J]. HEREDITAS(Beijing), 2015, 37(9): 899-910.

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多重环介导等温扩增技术研究进展

Progress in multiplex loop-mediated isothermal amplification technology

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