环介导等温扩增技术作为基因快速诊断方法的研究与展望

Research and Prospect of Loop-mediated Isothermal Amplification Assay as a Rapid Gene Screening Method

环介导等温扩增技术(LAMP)是一种新型分子诊断技术,具有灵敏度高、反应快速等优点,且仅利用简单的恒温水浴设备即可实现对特异性靶基因的高效扩增,在分子诊断领域具有巨大潜力。近几年来LAMP技术在分子诊断领域的研究非常广泛,涉及细菌、病毒、寄生虫的检测及食品转基因成分检验等诸多方面,受到越来越多学者认可,有研究者认为其可作为PCR的替补技术应用于临床基因诊断工作中。然而由于目前该技术缺乏规范化且成熟配套的上下游技术措施,如快速简单的核酸抽提技术、完善的应用体系和方案、防止交叉污染的有效措施及小型便携的检测设备等,因此在现场即时诊断领域的推广应用较为缓慢。为发挥利用LAMP技术在快速基因诊断方面的优越性,科研人员针对LAMP技术开发应用存在的问题开展了大量研究工作。立足于临床基因快速诊断的技术需求,对影响LAMP技术应用的核酸物质的快速抽提、LAMP检测体系的优化完善、结果判定、假阳性结果的规避、结果确诊及现场诊断一体化平台设计应用等领域的发展现状、需要攻克的难点及未来的应用展望进行综述,以期为未来的研究和开发指明方向,促进LAMP技术的不断完善及临床推广应用。

Loop-mediated isothermal amplification(LAMP)is a novel diagnostic technique requiring only simple isothermal heating device to efficiently amplify the targeted specific genes.Due to the high sensitivity and fastness,LAMP has showed to be a potential gene diagnostic tool since its discovery.In recent years,this technology is widely adopted in molecular diagnostics,involving the detection of bacteria,viruses,parasites,animal and plant GMO inspection and other food component determination.More and more researchers regard LAMP as an alternative method instead of PCR for clinical gene diagnosis.However,the lacking of normalized upstream and downstream technologies,such as fast nucleic acid extraction,ideal reaction systems,effective measures against cross contamination,portable detection devices and so on,leads to significant obstacles in its application for on-site diagnosis.To better meet the need of fast detection,many researches targeting to resolve the above associated problems have get primary results.In this article,we define LAMP as an early screening technique according to the demands of rapid clinical diagnosis.Further,we review the current research status and the limit of the related technologies involving fast nucleic acid extraction,optimized detection solutions,avoidance of false-positive results,re-check and determination of the results,the platform of field LAMP performance,and prospect the future technology development and application,aiming at highlighting the future research and development and promoting the constant improvement and clinical application of LAMP.