PDF(Pubmed)
Abstract：
The loop-mediated isothermal amplification (LAMP) technique is a great alternative to PCR-based methods, as it is fast, easy to use and works with high sensitivity and specificity without the need for expensive instruments. However, one of the limitations of LAMP is difficulty in achieving the simultaneous detection of several targets in a single tube, as the methodologies that allow this rely on fluorogenic probes containing specific target sequences, complicating their adaptation and the optimization of assays. Here, we summarize different methods for the development of multiplex LAMP assays based on sequence-specific detection, illustrated with a schematic representation of the technique, and evaluate their practical application based on the real-time detection and quantification of results, the possibility to visualize the results at a glance, the prior stabilization of reaction components, promoting the point-of-care use, the maximum number of specific targets amplified, and the validation of the technique in clinical samples. The various LAMP multiplexing methodologies differ in their operating conditions and mechanism. Each methodology has its advantages and disadvantages, and the choice among them will depend on specific application interests.
摘要：
环介导等温扩增（LAMP）技术是基于PCR的方法的绝佳替代方法，因为它很快，易于使用，具有高灵敏度和特异性，无需昂贵的仪器。然而,LAMP的局限性之一是难以实现在单个管中同时检测多个目标，作为允许这种方法依赖于含有特定靶序列的荧光探针的方法，使它们的适应和测定的优化复杂化。这里,我们总结了基于序列特异性检测的多重LAMP检测的不同方法，用技术的示意图来说明，并根据结果的实时检测和量化评估其实际应用，一目了然地可视化结果的可能性，反应组分的预先稳定，促进即时护理使用，扩增的特异性靶标的最大数量，以及该技术在临床样本中的验证。各种LAMP多路复用方法在其操作条件和机制方面不同。每种方法都有其优点和缺点，它们之间的选择将取决于特定的应用兴趣。
