Akhmetzianova Liana U, Mikhaylenko Constantin I, Chemeris Dmitry A, Khairitdinov Valery D, Sakhabutdinova Assol R, Gubaydullin Irek M, Garafutdinov Ravil R, Chemeris Alexey V
Institute of Biochemistry and Genetics, Ufa Federal Research Center, Russian Academy of Sciences, 71, prosp. Oktyabrya, Ufa 450054, Russia.
Department of Technology, Ufa State Petroleum Technological University, 1, st. Cosmonauts, Ufa 450064, Russia.
Biomolecules. 2025 May 9;15(5):690. doi: 10.3390/biom15050690.
Loop-mediated isothermal amplification (LAMP) was developed a quarter of a century ago, but it is still not exactly clear how this reaction proceeds. Only a few articles have focused on the kinetics of LAMP and the types of products formed. In this work, 10 types were identified and named. A basic dumbbell structure, Z6_dmb(1), consists of six zones and triggers the LAMP cycle. Due to self-priming, Z6_dmb(1) transforms into hairpin structure Z9_hp(1) and then into linearized strand Z9_li(1), carrying also strand Z6_dmb(2). Through similar transformations, it again generates strand Z6_dmb(1), completing the first LAMP cycle and starting a new one. The next stage of the exponential phase starts from two Z15_hp hairpin structures generated in the LAMP cycle, which next turn into Z15_li → Z27_hp → Z27_li → Z51_hp → and so forth. Modeling of a new type of the reaction, namely, pseudo-hemi-nested LAMP (phn-LAMP), was carried out. phn-LAMP involves three inner primers: two forward (FIP and extraFIP) and one backward inner primer, or vice versa. phn-LAMP has an advantage over LAMP involving loop or stem primers and over MIP-LAMP (multiple inner primers).
环介导等温扩增技术（LAMP）是在四分之一个世纪前开发的，但该反应具体是如何进行的仍不完全清楚。仅有少数文章关注LAMP的动力学以及所形成产物的类型。在这项研究中，鉴定并命名了10种类型。一种基本的哑铃结构Z6_dmb(1)由六个区域组成，并触发LAMP循环。由于自身引发，Z6_dmb(1)转变为发夹结构Z9_hp(1)，然后转变为线性化链Z9_li(1)，同时携带链Z6_dmb(2)。通过类似的转变，它再次生成链Z6_dmb(1)，完成第一个LAMP循环并开始新的循环。指数期的下一阶段从LAMP循环中产生的两个Z15_hp发夹结构开始，接着转变为Z15_li → Z27_hp → Z27_li → Z51_hp → 等等。对一种新型反应，即伪半巢式LAMP（phn-LAMP）进行了建模。phn-LAMP涉及三个内部引物：两个正向引物（FIP和extraFIP）和一个反向内部引物，反之亦然。phn-LAMP相对于涉及环引物或茎引物的LAMP以及MIP-LAMP（多个内部引物）具有优势。
