With the rapid development of isothermal amplification technology, DNA molecular diagnosis has become an important reference for clinical treatment. In this work, we have designed a DNA molecular diagnostic technology with LAMP-like sensitivity for nucleic acid analysis and detection based on only one pair of hairpin primers. This DNA molecular diagnostic technology consists of Bst DNA polymerase and one pair of hairpin primers, which are designed easily by adding a stem-loop structure to a target binding domain. When the target is present, the polymerization reaction between the hairpin primers and the target generates a specific dumbbell DNA similar to LAMP, which triggers cyclic amplification reactions to extend a series of long dsDNA products with repeated sequences by inserting fluorescent dye Eva Green observed the increase in fluorescence signal. In our method, using the hairpin primers-mediated isothermal polymerization amplification, we can specifically monitor 3-5 copies of the target nucleic acid in the system without labeling and temperature cycling in the reaction. In addition, serum samples from 13 patients with suspected schistosomiasis were targeted; we further demonstrated the ability of the technology to detect complex clinic samples, and its potentially inestimable applicability in clinic early molecular diagnostic research.

中文翻译：

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基于一对发夹引物介导的等温聚合扩增的具有LAMP样灵敏度的DNA分子诊断技术

随着等温扩增技术的快速发展，DNA分子诊断已成为临床治疗的重要参考。在这项工作中，我们设计了一种仅基于一对发夹引物进行核酸分析和检测的具有类似 LAMP 灵敏度的 DNA 分子诊断技术。这种 DNA 分子诊断技术由 Bst DNA 聚合酶和一对发夹引物组成，通过将茎环结构添加到目标结合域来轻松设计。当靶标存在时，发夹引物与靶标之间的聚合反应产生类似于LAMP的特异性哑铃DNA，通过插入荧光染料 Eva Green 触发循环扩增反应以扩展一系列具有重复序列的长 dsDNA 产物，观察到荧光信号的增加。在我们的方法中，使用发夹引物介导的等温聚合扩增，我们可以专门监测系统中目标核酸的 3-5 个拷贝，而无需在反应中进行标记和温度循环。此外，还针对13名疑似血吸虫病患者的血清样本；我们进一步证明了该技术检测复杂临床样本的能力，及其在临床早期分子诊断研究中潜在的不可估量的适用性。我们可以专门监测系统中3-5个拷贝的目标核酸，无需在反应中进行标记和温度循环。此外，还针对13名疑似血吸虫病患者的血清样本；我们进一步证明了该技术检测复杂临床样本的能力，及其在临床早期分子诊断研究中潜在的不可估量的适用性。我们可以专门监测系统中3-5个拷贝的目标核酸，无需在反应中进行标记和温度循环。此外，还针对13名疑似血吸虫病患者的血清样本；我们进一步证明了该技术检测复杂临床样本的能力，及其在临床早期分子诊断研究中潜在的不可估量的适用性。