Yan Xiaoyu, Tang Min, Yang Jianru, Diao Wei, Ma Hongmin, Cheng Wenbin, Que Haiying, Wang Tong, Yan Yurong
Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), College of Laboratory Medicine, Chongqing Medical University Chongqing 400016 China
Department of Clinical Laboratory, Affiliated Hospital of Zunyi Medical University Zunyi 563003 China.
RSC Adv. 2018 Sep 12;8(55):31710-31716. doi: 10.1039/c8ra06480f. eCollection 2018 Sep 5.
Sensitive and specific detection of HIV-related DNA is of great importance for early accurate diagnosis and therapy of HIV-infected patients. Here, we developed a one-step and rapid fluorescence strategy for HIV-related DNA detection based on strand displacement amplification and a Mg-dependent DNAzyme reaction. In the presence of target HIV DNA, it can hybridize with template DNA and activate strand displacement amplification to generate numerous DNAzyme sequences. With the introduction of Mg, DNAzyme can be activated to circularly cleave the substrate DNA, which leads to the separation of fluorophore reporters from the quenchers, resulting in the recovery of the fluorescence. Under the optimal experimental conditions, the established biosensing method can detect target DNA down to 61 fM with a linear range from 100 fM to 1 nM, and discriminate target DNA from mismatched DNA perfectly. In addition, the developed biosensing strategy was successfully applied to assay target DNA spiked into human serum samples. With the advantages of fast, easy operation and high-performance, this biosensing strategy might be an alternative tool for clinical diagnosis of HIV infection.
灵敏且特异的HIV相关DNA检测对于HIV感染患者的早期准确诊断和治疗至关重要。在此，我们基于链置换扩增和镁依赖的脱氧核酶反应，开发了一种用于HIV相关DNA检测的一步快速荧光策略。在目标HIV DNA存在的情况下，它可与模板DNA杂交并激活链置换扩增，以产生大量脱氧核酶序列。随着镁的引入，脱氧核酶可被激活以循环切割底物DNA，这导致荧光团报告分子与淬灭剂分离，从而使荧光恢复。在最佳实验条件下，所建立的生物传感方法能够检测低至61 fM的目标DNA，线性范围为100 fM至1 nM，并且能够完美地区分目标DNA与错配DNA。此外，所开发的生物传感策略成功应用于检测加标到人类血清样本中的目标DNA。凭借快速、操作简便和高性能的优点，这种生物传感策略可能成为HIV感染临床诊断的一种替代工具。
