环介导等温扩增技术的研究进展

Research progress of loop-mediated isothermal amplification technology

环介导等温扩增(loop-mediated isothermal amplification,LAMP)技术是一种新型的恒温核酸扩增技术。与传统的PCR技术相比,该技术能在恒定温度65℃条件下,利用4条特异性设计的引物,2条内引物(forward inner primer,FIP)、(backward inner primer,BIP)和2条外引物F3 primer、B3 primer和一种具有链置换特性的Bst.DNA聚合酶。在30~60 min内对目的DNA序列进行快速、高效扩增,其扩增产物为大量的双链DNA。通过对产物的处理,结合双链DNA特点,根据目标DNA的含量直接或者间接得到待测物含量,从而达到一个较高的灵敏度检测。利用该技术实现了在多种领域,包括食品致病菌检测,食品微生物含量检测以及生物医学等的广泛应用。本文主要对LAMP的技术原理、产物检测方法、应用领域、与其它方法比较的优缺点以及发展现状进行相关的论述。

Loop-mediated isothermal amplification(LAMP)is a novel thermostatic nucleic acid amplification technique.Compared with the traditional polymerase chain reaction(PCR)technology,LAMP can amplify the target DNA at 65 ℃ by using 4 primers,2 inner primers,FIP(forward inner primer)and(backward inner primer,B1 P),2 outer primers,F3 primer(forward outer primer)and B3 primer(backward outer primer)designed specifically,and the Bst.DNA polymerase.The target DNA was rapidly and efficiently amplified within 30-60 min and the amplified products were a large number of double-stranded DNA(dsDNA).Through processing and combining the characteristics of the dsDNA,the content of the analyte will be directly or indirectly obtained.Therefore,it can achieve a highly sensitivity detection.LAMP has a wide range of application in various fields,including the detection of food pathogens food microbial content and the wide application of biomedicine.This work mainly discussed the technical principle,product detection methods,application fields,advantages and disadvantages compared with other methods and development status of LAMP.