Trypsin-Amplified Aerolysin Nanopore Amplified Sandwich Assay for Attomolar Nucleic Acid and Single Bacteria Detection
Trypsin-Amplified Aerolysin Nanopore Amplified Sandwich Assay for Attomolar Nucleic Acid and Single Bacteria Detection
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用于阿摩尔核酸和单一细菌检测的胰蛋白酶放大气溶素纳米孔放大夹心法
DOI：
10.1021/acs.analchem.9b03717
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发表时间：
2019-11-05
影响因子：
7.4
通讯作者：
Li, Jinghong
中科院分区：
文献类型：
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作者：
Son, Fengge;Deng, Ruijie;Li, Jinghong
纳米孔技术有望用于下一代基于核酸的诊断。然而，在低浓度下仍然很难实现序列保留。在此，我们提出了一种胰蛋白酶激活的催化反应放大检测，这大大提高了纳米孔技术的灵敏度。基于核酸探针设计，所提出的胰蛋白酶扩增的纳米孔扩增的夹心测定（tNASA）可以有助于高出约100 000倍的灵敏度。值得注意的是，tNASA能够通过使用小型化的电流放大器而无需比对算法来进行attomolar核酸和单细胞检测。它还允许血清中的10种病原物种被准确和稳健地分析，从而用于传染病的诊断。tNASA可能会发展用于核酸检测的纳米孔技术的构建，并将促进其转化为口袋诊断和精确医学。
Nanopore technology is promising for the next-generation of nucleic acid based diagnosis. However, sequence reservation could still be hardly achieved in low-concentration. Herein, we propose a trypsin-activated catalysis reaction for amplified detection, which substantially improves the sensitivity of nanopore technique. The proposed trypsin-amplified nanopore amplified sandwich assay (tNASA) could contribute to a sensitivity approximately 100 000 times higher based on nucleic acid probe design. Remarkably, tNASA is capable of attomolar nucleic acid and single cell detection by using a miniaturized current amplifier without alignment algorithm. Also it allows 10 pathogenic species in serum to be accurately and robustly profiled, thus be utilized for the diagnosis of infectious diseases. tNASA may evolve the construction of nanopore techniques for nucleic acid detection and would facilitate its translation for pocket diagnosis and precision medicine.