Loop-mediated isothermal amplification (LAMP) is the most popular technique to amplify nucleic acid sequence without the use of temperature cycling. However, LAMP is often confounded by false-positive results, arising from interactions between (hetero-dimer) or within (self-dimerization) primers, off-target hybrids and carryover contaminants. Here, we devised a new LAMP technique that is self-avoiding molecular recognition system (SAMRS) components and antarctic thermal sensitive uracil-DNA-glycosylase (AUDG) enzyme-assisted, termed AUDG-SAMRS-LAMP. Incorporating SAMRS components into 3'-ends of LAMP primers can improve assay's specificity, which completely prevents the non-specific amplification yielding from off-target hybrids and undesired interactions between or within primers. Adding AUDG into reaction mixtures can effectively eliminate the false-positive results arising from carryover contamination, thus the genuine positive reactions are generated from the amplification of target templates. Furthermore, AUDG-SAMRS-LAMP results are confirmed using a new analysis strategy, which is developed for detecting LAMP amplicons by lateral flow biosensor (LFB). Only a single labeled primer is required in the analysis system, thus the false positive results arising from hybridization (the labeled primer and probe, or between two labeled primers) are avoided. Hence, the SAMRS components, AUDG and LFB convert traditional LAMP from a technique suited for the research laboratory into one that has practical value in the field of diagnosis. Human Tuberculosis (TB) is caused by infection with members of Mycobacterium tuberculosis complex (MTC), which are detected by the AUDG-SAMRS-LAMP technique to demonstrate the availability of target analysis. The proof-of-concept method can be reconfigured to detect various nucleic acids by redesigning the specific primers.

中文翻译：

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环介导等温扩增，使用自回避分子识别系统和南极热敏尿嘧啶-DNA-糖基化酶检测核酸，防止携带污染

环介导等温扩增 (LAMP) 是最流行的核酸序列扩增技术，无需使用温度循环。然而，LAMP 经常被假阳性结果所混淆，假阳性结果是由（异二聚体）之间或（自二聚化）引物、脱靶杂交和残留污染物之间的相互作用引起的。在这里，我们设计了一种新的 LAMP 技术，它是自回避分子识别系统 (SAMRS) 组件和南极热敏尿嘧啶-DNA-糖基化酶 (AUDG) 酶辅助，称为 AUDG-SAMRS-LAMP。将 SAMRS 组分加入 LAMP 引物的 3' 端可以提高检测的特异性，这完全防止了脱靶杂交产生的非特异性扩增以及引物之间或内部的不希望的相互作用。在反应混合物中加入 AUDG 可以有效消除残留污染引起的假阳性结果，从而产生真正的阳性反应来自目标模板的扩增。此外，AUDG-SAMRS-LAMP 结果使用一种新的分析策略得到确认，该策略是为通过侧流生物传感器 (LFB) 检测 LAMP 扩增子而开发的。分析系统只需要一个标记引物，避免了杂交（标记引物和探针，或两个标记引物之间）产生的假阳性结果。因此，SAMRS 组件 AUDG 和 LFB 将传统 LAMP 从一种适合研究实验室的技术转变为一种在诊断领域具有实用价值的技术。人类结核病 (TB) 是由结核分枝杆菌复合群 (MTC) 成员感染引起的，通过 AUDG-SAMRS-LAMP 技术检测到这些成员，以证明目标分析的可用性。通过重新设计特定引物，可以重新配置概念验证方法以检测各种核酸。