One-tube smart genetic testing via coupling isothermal amplification and three-way nucleic acid circuit to glucometers
One-tube smart genetic testing via coupling isothermal amplification and three-way nucleic acid circuit to glucometers
复制标题
通过将等温扩增和三路核酸电路与血糖仪耦合的一管智能基因测试
DOI：
10.1016/j.aca.2020.01.068
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发表时间：
2020-04-15
影响因子：
6.2
通讯作者：
Li, Bingling
中科院分区：
文献类型：
--
作者：
Guo, Lulu;Lu, Baiyang;Li, Bingling
对便携式诊断的迫切需求促进了一种新的传感策略，该策略使用个人血糖仪（PGM）来检测非葡萄糖目标。尽管在目标范围和传感原理方面取得了很大进展，但最终信号背景比低和难以实现单管智能分析等问题仍然存在，并挑战了基因检测的实际应用。在这里，我们提出了一个实际的解决方案，通过耦合等温扩增（即LAMP）和三路可重复催化发夹组装（即CHA）的PGM。它允许从基因组信息直接转换到商业便携式设备，具有超高灵敏度，特异性和增强的信噪比。与以前报道的无信号放大相比，CHA的引入成功地将信号幅度提高了至少12.5倍。更重要的是，通过引入有效的基于三向连接的转导，我们还创新性地开发了基于PGM检测的单管逻辑或多重分析策略。最终的PGM信号可以直接读出坂崎克罗伊氏菌（ompA）和大肠杆菌（malB）中两种食源性细菌基因的4种情况，最低检测量可达100个分子拷贝（6.6 × 10（-18）M）以下。据信，这种LAMP-CHA-PGM方法已经是灵敏的、特异的，并且对于实际便携式基因诊断具有巨大潜力。(C)2020 Elsevier B. V.保留所有权利。
Urgent demand for portable diagnosis has promoted a new sensing strategy that uses personal glucometer (PGM) to detect non-glucose targets. Even though great progresses have been achieved in terms of target range and sensing principle, issues such as low final signal-to-background ratio and hard-to-realize one-tube smart analysis still exist and challenge real-world applications in gene detection. Here we propose a practical solution via coupling isothermal amplification (i.e. LAMP) and three-way amplifiable catalytic hairpin assembly (i.e. CHA) to a PGM. It allows direct transduction from genomic information to commercial portable devices with all of ultra-high sensitivity, specificity and enhanced signal-to-noise ratio. Compared with previous report without signal amplification, the introduction of CHA has successfully improved the signal amplitude by at least 12.5 folds. More importantly, through importing an effective three-way junction based transduction, we also innovatively develop a one-tube logical or multiplex analysis strategy in PGM based detection. Totally four situations of two foodborne bacteria genes, in Cronobacter sakazakii (ompA) and Escherichia coli (malB), could be directly readout using the final PGM signals, with the lowest detection amount down to less than 100 molecular copies (6.6 x 10(-18) M). It is believed such a LAMP-CHA-PGM method has been already sensitive, specific, and of great potential for practically portable gene diagnostics. (C) 2020 Elsevier B.V. All rights reserved.