State Key Lab of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, Jilin, 130022, China.
University of Science and Technology of China, Hefei, Anhui 230026, China.
Angew Chem Int Ed Engl. 2022 Oct 4;61(40):e202209496. doi: 10.1002/anie.202209496. Epub 2022 Aug 25.
Sensitive, rapid and low-cost nucleic acid detection is critical for controlling infectious pathogens. Here, we develop a ready-to-use and multimodal detection based on a rebuilding-free, ultrasensitive and selective strategy named dual hairpin ligation-induced isothermal amplification pro (DHLApro). Taking influenza A, influenza B, MERS-CoV, SARS-CoV-2 as model targets, we demonstrate DHLApro provides ≈zM level ultra-sensitivity, being equaling to 0.45 copy/μL in original sample. By simply changing the recognition module, a set of DHLApro components can be applied to a new target without performance loss. Moreover, DHLApro innovatively allows flexible logic/multiplex assay using one set of primer, for example, the "N pathogens-in-1" OR gate screening and accurate multi-channel multiplex assay. Compared with traditional methods, the cost of this logic/multiplex assay has been largely reduced and the cross-interference between the multiple primer sets is also avoided.
灵敏、快速且低成本的核酸检测对于控制传染病病原体至关重要。在这里，我们开发了一种基于免重建、超灵敏和选择性策略的即用型多模态检测方法，称为双发夹连接诱导等温扩增探针 (DHLApro)。以甲型流感、乙型流感、中东呼吸综合征冠状病毒、严重急性呼吸综合征冠状病毒 2 型为模型靶标，我们证明了 DHLApro 提供了 ≈zM 级别的超灵敏性，在原始样本中达到了 0.45 拷贝/μL。通过简单地改变识别模块，一组 DHLApro 组件可以应用于新的靶标而不会降低性能。此外，DHLApro 创新性地允许使用一组引物进行灵活的逻辑/多重分析，例如，"N 种病原体中的 1 种"或门筛选和准确的多通道多重分析。与传统方法相比，这种逻辑/多重分析的成本大大降低，并且避免了多个引物组之间的交叉干扰。
