环介导等温扩增,LAMP,是目前最流行的等温核酸扩增技术,因此,几个商业,现成的主混音蓬勃发展。不幸的是,确定他们表现的独立研究是有限的。本研究对现有的即用型商用LAMP主混合物WarmStart®LAMP套件进行了独立评估,LavaLAMP™DNAMasterMix,SaphirBstTurboGreenMaster,OptiGeneFastMasterMixISO-004和SynLAMPMix。为了减少偏见,三种不同的基因,即ttr(沙门氏菌。),rfbE(E.大肠杆菌O157),和hly(单核细胞增生李斯特菌),有针对性。比较是基于扩增速度,随着DNA浓度的降低,以及五种典型的LAMP反应添加剂(甜菜碱,DMSO,普鲁兰,TMAC,和GuHCl)。在不同的主混合物中观察到显著差异。OptiGene提供最快的扩增,并显示与评估的补充剂相关的有害影响较小。在测试的化学物质中,支链淀粉在扩增速度方面提供了最好的结果。值得注意的是,不同的添加剂对母料混合物的影响不同。总的来说,当前的研究提供了对商用LAMP主混合物性能的见解,这对于科学界在开发新方法时更好地选择合适的试剂具有价值。
Loop-mediated isothermal amplification,
LAMP, is nowadays the most popular isothermal nucleic acid amplification technique, and as such, several commercial, ready-to-use master mixes have flourished. Unfortunately, independent studies to determine their performance are limited. The current study performed an independent evaluation of the existing ready-to-use commercial
LAMP master mixes WarmStart®
LAMP Kit, LavaLAMP™ DNA Master Mix, Saphir Bst Turbo GreenMaster, OptiGene Fast Master Mix ISO-004, and SynLAMP Mix. To reduce bias, three different genes, namely ttr (Salmonella spp.), rfbE (E. coli O157), and hly (Listeria monocytogenes), were targeted. The comparison was based on amplification speed, performance with decreasing DNA concentrations, and the effect of five typical
LAMP reaction additives (betaine, DMSO, pullulan, TMAC, and GuHCl). Significant differences were observed among the different master mixes. OptiGene provided the fastest amplification and showed less detrimental effects associated with the supplements evaluated. Out of the chemicals tested, pullulan provided the best results in terms of amplification speed. It is noteworthy that the different additives impacted the master mixes differently. Overall, the current study provides insights into the performance of commercial
LAMP master mixes, which can be of value for the scientific community to better select appropriate reagents when developing new methods.