A signal-flexible gene diagnostic strategy coupling loop-mediated isothermal amplification with hybridization chain reaction
A signal-flexible gene diagnostic strategy coupling loop-mediated isothermal amplification with hybridization chain reaction
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一种信号灵活的基因诊断策略，将环介导的等温扩增与杂交链式反应相结合
DOI：
10.1016/j.aca.2019.06.048
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发表时间：
2019-11-04
影响因子：
6.2
通讯作者：
Du, Yan
中科院分区：
文献类型：
--
作者：
Dong, Qing;Liu, Quanyi;Du, Yan
近年来的研究表明，环介导等温核酸扩增（LAMP）技术与一步链置换（OSD）技术相结合，有助于提高基因检测的序列特异性。然而，由于OSD不能进行信号放大，因此信噪比或可观察到的信号变化通常不足以满足实际使用。为了克服这一挑战，本文开发了一种更先进和实用的传感原理，其中OSD被可重复的核酸电路，杂交链反应（HCR）所取代。采用高度传染性诺如病毒（NoV）作为模型靶标。与LAMP-OSD相比，LAMP-HCR可在2%的粪便样品中检测到30个拷贝的NoV基因，且信号变化和信号背景比显著增大。因此，实现了更可靠的检测。此外，由于HCR的高兼容性，最终的LAMP-HCR产品可以灵活地转换为不同类型的读数，包括荧光，流式细胞仪（FCM），甚至个人血糖仪（PGM）。这进一步扩大了检测的操作环境，从医院实验室、床边到当地药店的潜在现成设备。特别是当使用FCM或PGM时，在磁珠（MB）的辅助下，检测显示出对复杂生物基质的更高耐受能力。(C)2019 Elsevier B. V.版权所有。
Recent study proves that the combination of loop mediated isothermal nucleic acid amplification (LAMP) with one-step strand displacement (OSD) is of great help to improve the sequence specificity during genetic detection. However, because OSD is incapable of signal amplification, the signal-to-noise ratio or the observable signal change may be usually not significant enough to satisfy practical usage. With the purpose to overcome this challenge, herein a more advanced and practical sensing principle is developed with the OSD replaced by an amplifiable nucleic acid circuit, hybridization chain reaction (HCR). The very contagious norovirus (NoV) was employed as the model target. Compared with LAMP-OSD, the LAMP-HCR can detect as few as 30 copies of NoV gene in 2% fecal samples with significantly enlarged signal change and signal-to-background ratio. Therefore, more reliable detection is achieved. Moreover, due to the high compatibility of HCR, the final LAMP-HCR products can be flexibly transduced into different types of readouts, including fluorescence, flow cytometer (FCM) and even a personal glucose meter (PGM). This further enlarges the operating environments for the detection from hospital labs, bedsides, to potential off-the-shelf devices in local pharmacies. Especially when using FCM or PGM, with the assistance of magnetic beads (MBs), the detection shows even higher tolerance capability to complicated biological matrices. (C) 2019 Elsevier B.V. All rights reserved.