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Programmable hybridization assemble nicked displacement amplification for detecting ricin toxin
Sensors and Actuators B: Chemical ( IF 8.4 ) Pub Date : 2022-06-11 , DOI: 10.1016/j.snb.2022.132139
Yu Wang, Yuan Peng, Jialei Bai, Shuang Li, Dianpeng Han, Shuyue Ren, Kang Qin, Sen Li, Tie Han, Huanying Zhou, Zhixian Gao

Isothermal amplification, such as hybridization chain reaction (HCR), is a simple and reliable method for detecting signal amplification. However, the hairpin in HCR will not fully participate in the reaction. And after the hairpin is opened, the distance between the fluorophore and the quencher does not change much. Therefore, the signal magnification is limited. Here, we designed a new isothermal amplification method named hybridization assemble nicked displacement amplification (HANDA), combining HCR and strand displacement amplification (SDA) ingeniously. HANDA first triggers HCR through the target sequence to form long double-strand DNA (dsDNA) with gaps. Then SDA is performed from the gap to obtain a large amount of single-stranded DNA (ssDNA), so as to achieve the purpose of double signal amplification. The base sequence of DNA hairpin had also been optimized. The best sequence design rule was found and had universal applicability. We have demonstrated that HANDA combined with DNA barcodes can be used for trace detection of ricin. This new isothermal amplification method provides an effective and universal platform for the trace detection of various toxic substances.



中文翻译:

用于检测蓖麻毒素的可编程杂交组装缺口置换扩增

等温扩增,如杂交链式反应 (HCR),是一种简单可靠的信号放大检测方法。但是,HCR 中的发夹不会完全参与反应。并且发夹打开后,荧光团和猝灭剂之间的距离变化不大。因此,信号放大倍数受到限制。在这里,我们设计了一种新的等温扩增方法,称为杂交组装缺口置换扩增(HANDA),将HCR和链置换扩增(SDA)巧妙地结合起来。HANDA首先通过靶序列触发HCR,形成带缺口的长双链DNA(dsDNA)。然后从缺口处进行SDA,获得大量的单链DNA(ssDNA),从而达到双重信号放大的目的。DNA发夹的碱基序列也进行了优化。找到了最佳的序列设计规则并具有普遍适用性。我们已经证明 HANDA 结合 DNA 条形码可用于蓖麻毒素的痕量检测。这种新的等温扩增方法为各种有毒物质的痕量检测提供了一个有效且通用的平台。

更新日期:2022-06-14
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