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首页> 外文期刊>RSC Advances >Detection of nucleic acids via G- quadruplexcontrolled L- cysteine oxidation and catalyzed hairpin assembly- assisted signal amplification
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Detection of nucleic acids via G- quadruplexcontrolled L- cysteine oxidation and catalyzed hairpin assembly- assisted signal amplification

机译：通过G-四逆转录的L-半胱氨酸氧化和催化发夹组装辅助信号放大检测核酸

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摘要

The development of simple, sensitive and cost-effective methods for specific nucleic acid detection has attracted tremendous attention due to its importance to the early diagnosis of genetic diseases and to biodefense applications. In this work, we demonstrated a fluorescent turn-off mode DNA assay based on l-cysteine-modulated synthesis of CdTe quantum dots (CdTe QDs), horseradish peroxidase-mimicking G-quadruplex-hemin-K+ complex controlled oxidation of l-cysteine to cystine, and catalyzed hairpin assembly (CHA)-assisted signal amplification. After the addition of target DNA, the CHA signal amplification reaction was triggered and numerous H1-H2 double-stranded DNA were formed, initiating the release of G-quadruplex sequences in H2 simultaneously. Thus, the degree of inhibition of the synthesis of CdTe QDs is proportional to the concentration of the G-quadruplex sequence in this method. In contrast, when the target DNA was absent, the CHA could not be triggered, and the fluorescence signal was high due to the remaining intact l-cysteine. Under optimal experimental conditions, the homogeneous fluorescence method achieved the detection of HIV DNA with a linear range from 0.1 pM to 1 nM and a detection limit of 0.12 pM. This novel biosensor exhibits excellent specificity in differentiating DNA sequences with a single-base and two-base mismatch. To the best of our knowledge, this a label-free and highly sensitive bioassay utilizing CHA-assisted signal amplification and G-quadruplex control of in situ synthesis of CdTe QDs strategy was not reported in previous. Thus, this proposed strategy is anticipated to find use in basic biochemical research and clinical diagnosis.
机译：由于其对遗传疾病早期诊断和生物义应用的重要性，对特定核酸检测的简单，敏感和经济高效的方法的发展引起了巨大的关注。在这项工作中，我们证明了基于L-半胱氨酸调节的CDTE量子点（CDTE QD）的荧光关断模式DNA测定，辣根过氧化物酶模拟G-Quadreple--Hemin-K +复合控制氧化L-半胱氨酸至胱氨酸，催化发夹组件（CHA）译力信号放大。在添加靶DNA之后，触发CHA信号扩增反应，并形成许多H1-H2双链DNA，同时在H 2中引发G-Quadwlex序列的释放。因此，CDTE QD的合成的抑制程度与该方法中的G- Quadflex序列的浓度成比例。相反，当不存在靶DNA时，不能触发CHA，并且由于剩余的完整L-半胱氨酸，荧光信号很高。在最佳实验条件下，均匀荧光方法达到了从0.1pm至1nm的线性范围检测HIV DNA，检测限为0.12μm。这种新的生物传感器在用单碱基和双碱错配的区分DNA序列中表现出优异的特异性。据我们所知，这种无标记且高度敏感的生物测定，利用CHA辅助信号放大和G-QUADRUPLEX对原位合成CDTE QDS策略的控制，在之前没有报道。因此，预计这一拟议的策略将在基础生化研究和临床诊断中找到使用。

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  • 来源
    《RSC Advances》 |2018年第71期|共6页
  • 作者

    Chen Piaopiao; Hu Pingyue; Huang Ke; Sawyer Erica; Sun Ke; Ying Binwu; Wei Xiawei; Geng Jia;

  • 作者单位

    Sichuan Univ West China Hosp State Key Lab Biotherapy Dept Lab Med Chengdu 610041 Sichuan Peoples R China;

    Sichuan Normal Univ Coll Chem &

    Mat Sci Chengdu 610068 Sichuan Peoples R China;

    Sichuan Normal Univ Coll Chem &

    Mat Sci Chengdu 610068 Sichuan Peoples R China;

    Sichuan Univ West China Hosp State Key Lab Biotherapy Dept Lab Med Chengdu 610041 Sichuan Peoples R China;

    Sichuan Univ West China Hosp State Key Lab Biotherapy Dept Lab Med Chengdu 610041 Sichuan Peoples R China;

    Sichuan Univ West China Hosp State Key Lab Biotherapy Dept Lab Med Chengdu 610041 Sichuan Peoples R China;

    Sichuan Univ West China Hosp State Key Lab Biotherapy Dept Lab Med Chengdu 610041 Sichuan Peoples R China;

    Sichuan Univ West China Hosp State Key Lab Biotherapy Dept Lab Med Chengdu 610041 Sichuan Peoples R China;

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