基于富G序列的无标记荧光传感器检测单链核酸

引用本文: 任林娇, 薛梦晓, 陈青华, 魏铭航, 张培, 秦自瑞, 闫艳霞, 姜利英. 基于富G序列的无标记荧光传感器检测单链核酸[J]. 分析化学, 2023, 51(6): 962-971. doi: 10.19756/j.issn.0253-3820.221546 shu

Citation: REN Lin-Jiao, XUE Meng-Xiao, CHEN Qing-Hua, WEI Ming-Hang, ZHANG Pei, QIN Zi-Rui, YAN Yan-Xia, JIANG Li-Ying. Label-free Fluorescent Sensor for Detection of Single-stranded Nucleic Acids Based on G-rich Sequence[J]. Chinese Journal of Analytical Chemistry, 2023, 51(6): 962-971. doi: 10.19756/j.issn.0253-3820.221546 shu

基于富G序列的无标记荧光传感器检测单链核酸

任林娇 ^1, ,
薛梦晓 ^1, ,
陈青华 ^{1,
,} ,
魏铭航 ^1, ,
张培 ^1, ,
秦自瑞 ^1, ,
闫艳霞 ^2, ,
姜利英 ^{1,3,
,}

1.
郑州轻工业大学电气信息工程学院, 郑州 450002;
2.
郑州轻工业大学计算机与通信工程学院, 郑州 450002;
3.
郑州轻工业大学量子科技研究院, 郑州 450002

通讯作者: 陈青华,E-mail:2006007@zzuli.edu.cn; 姜利英,E-mail:jiangliying@zzuli.edu.cn

基金项目:
国家自然科学基金项目(Nos.62073299,12004348,61801436)、中原科技创新领军人才项目(No.224200510026)和河南省科技攻关项目(Nos.212102210196,212102210002,212102210278,212102210008)资助。

摘要: 具有特定构象的富G序列（如G-四链体和G-三链体）与荧光染料相互作用可有效增强其荧光信号强度，被广泛应用于构建无标记荧光生物传感。本研究以硫磺素T（Thioflavin T，ThT）为荧光染料，构建了两种基于富G序列的无标记荧光传感器，用于检测阿尔兹海默病标志物β-淀粉样蛋白（β-Amyloid protein，Aβ）的基因序列。实验结果表明，分子发夹茎长为4个碱基时，富G序列以G-三链体的结构存在，以此构建的G-三链体传感器的输出信号随Aβ基因浓度增加而降低，线性检测范围为1~100 nmol/L，检出限为0.3 nmol/L（S/N=3）。分子发夹茎长为8个碱基且5'端添加碱基AATT时，其与Aβ基因结合后，富G序列多以G-四链体结构存在，以此构建的G-四链体传感器的输出信号随Aβ基因浓度增加而增强，线性检测范围为0.1~100 nmol/L，检出限为0.04 nmol/L （S/N=3）。两种传感器制备过程相似但检测原理不同，为富G序列的深入研究与应用提供了参考，同时为单链核酸的无标记荧光检测提供了新的思路。

关键词:

English

Label-free Fluorescent Sensor for Detection of Single-stranded Nucleic Acids Based on G-rich Sequence

1.
School of Electrical and Information Engineering, Zhengzhou University of Light Industry, Zhengzhou 450002, China;
2.
College of Computer and Communication Engineering, Zhengzhou University of Light Industry, Zhengzhou 450002, China;
3.
Academy for Quantum Science and Technology, Zhengzhou University of Light Industry, Zhengzhou 450002, China

Corresponding author: CHEN Qing-Hua, ; JIANG Li-Ying,

Received Date: 03 November 2022
Revised Date: 03 April 2023
Fund Project:
Supported by the National Natural Science Foundation of China (Nos. 62073299, 12004348, 61801436), the Project of Central Plains Science and Technology Innovation Leading Talents (No. 224200510026) and the Scientific and Technological Projects in Henan Province, China (Nos. 212102210196, 212102210002, 12102210278, 212102210008).

Abstract: G-rich sequences with specific conformations (e.g. G-quadruplex and G-triplex) can interact with fluorescent dyes to enhance their fluorescence signal intensity, and are widely used for label-free fluorescent biosensing. In this study, two label-free fluorescent sensors based on G-rich sequences were constructed for detection of the gene sequence of β-amyloid protein (Aβ), a marker of Alzheimer's syndrome, using thiosemicarbazone T (ThT) as the fluorescent dye. The experimental results showed that the G-rich sequences were present as G-triplexes when the length of the hairpin stem was 4 base pairs, and the output signal of the G-triplex sensor decreased with increasing concentration of Aβ gene, with a linear detection range of 1-100 nmol/L and a detection limit of 0.3 nmol/L (S/N = 3). When the length of the hairpin stem was 8 base pairs and the base AATT was added at the 5' end, the G-rich signal decreased with the concentration of Aβ gene. The G-rich sequences were mostly present as G-quadruplexes after binding to Aβ gene, and the output signal of the G-quadruplex sensor was enhanced with increasing concentration of Aβ gene, with a linear detection range of 0.1-100 nmol/L and a detection limit of 0.04 nmol/L (S/N = 3). The two sensors were prepared in a similar process but with different detection principles, providing a basis for further research and application of G-rich sequences. It also provided a new idea for the label-free fluorescence detection of single-stranded nucleic acids.

Key words:

1. [1]
  ZHANG N, WANG L, DENG X, LIANG R, SU M, HE C, HU L, SU Y, REN J, YU F, DU L, JIANG S. J. Med. Virol., 2020, 92(4):408-417.ZHANG N, WANG L, DENG X, LIANG R, SU M, HE C, HU L, SU Y, REN J, YU F, DU L, JIANG S. J. Med. Virol., 2020, 92(4):408-417.
2. [2]
  LI L, LI S, WU N, WU J, WANG G, ZHAO G, WANG J. ACS Synth. Biol., 2019, 8(10):2228-2237.LI L, LI S, WU N, WU J, WANG G, ZHAO G, WANG J. ACS Synth. Biol., 2019, 8(10):2228-2237.
3. [3]
  CHOI J H, LIM J, SHIN M, PAEK S H, CHOI J W. Nano Lett., 2021, 21(1):693-699.CHOI J H, LIM J, SHIN M, PAEK S H, CHOI J W. Nano Lett., 2021, 21(1):693-699.
4. [4]
  YANG C, CHU X, ZENG L, RHOUATI A, ABBAS F, CUI S, LIN D. Biosensors, 2022, 12(6):423.YANG C, CHU X, ZENG L, RHOUATI A, ABBAS F, CUI S, LIN D. Biosensors, 2022, 12(6):423.
5. [5]
  STOBIECKA M, CHAŁUPA A. Chem. Pap., 2015, 69(1):62-76.STOBIECKA M, CHAŁUPA A. Chem. Pap., 2015, 69(1):62-76.
6. [6]
  LIU Zhuo-Liang, TAO Cheng-An, WANG Jian-Fang. Chin. J. Anal. Chem., 2020, 48(2):153-163. 刘卓靓, 陶呈安, 王建方. 分析化学, 2020, 48(2):153-163.
7. [7]
  XIONG Y, DAI J, ZHANG Y, ZHOU C, YUAN H, XIAO D. Anal. Methods, 2021, 13(21):2391-2395.XIONG Y, DAI J, ZHANG Y, ZHOU C, YUAN H, XIAO D. Anal. Methods, 2021, 13(21):2391-2395.
8. [8]
  WANG K, ZHAI F H, HE M Q, WANG J, YU Y L, HE R H. Anal. Bioanal. Chem., 2019, 411(19):4569-4576.WANG K, ZHAI F H, HE M Q, WANG J, YU Y L, HE R H. Anal. Bioanal. Chem., 2019, 411(19):4569-4576.
9. [9]
  MA G, YU Z, ZHOU W, LI Y, FAN L, LI X. J. Phys. Chem. B, 2019, 123(26):5405-5411.MA G, YU Z, ZHOU W, LI Y, FAN L, LI X. J. Phys. Chem. B, 2019, 123(26):5405-5411.
10. [10]
  SHAHSAVAR K, SHOKRI E, HOSSEINI M. Microchem. J., 2020, 158:105277.SHAHSAVAR K, SHOKRI E, HOSSEINI M. Microchem. J., 2020, 158:105277.
11. [11]
  LI Z B, ZOU S Y, WU S J, MIAO X M, MA D L. Talanta, 2020, 221:121661.LI Z B, ZOU S Y, WU S J, MIAO X M, MA D L. Talanta, 2020, 221:121661.
12. [12]
  CHEN N, LI J, FENG X, YANG Y, ZHU L, CHEN X, LIU X, LI Y, WANG C, XIA L. Microchim. Acta, 2020, 187(8):432.CHEN N, LI J, FENG X, YANG Y, ZHU L, CHEN X, LIU X, LI Y, WANG C, XIA L. Microchim. Acta, 2020, 187(8):432.
13. [13]
  WANG S, SUN J, ZHAO J, LU S, YANG X. Anal. Chem., 2018, 90(5):3437-3442.WANG S, SUN J, ZHAO J, LU S, YANG X. Anal. Chem., 2018, 90(5):3437-3442.
14. [14]
  LU S, WANG S, ZHAO J, SUN J, YANG X. Anal. Chem., 2017, 89(16):8429-8436.LU S, WANG S, ZHAO J, SUN J, YANG X. Anal. Chem., 2017, 89(16):8429-8436.
15. [15]
  WU Z F, ZHOU H, HE J, LI M, MA X M, XUE J, LI X, FAN X L. Analyst, 2019, 144(17):5201-5206.WU Z F, ZHOU H, HE J, LI M, MA X M, XUE J, LI X, FAN X L. Analyst, 2019, 144(17):5201-5206.
16. [16]
  QIN S, CHEN X, XU Z, LI T, ZHAO S, HU R, ZHU J, LI Y, YANG Y, LIU M. Anal. Bioanal. Chem., 2022, 414(20):6149- 6156.QIN S, CHEN X, XU Z, LI T, ZHAO S, HU R, ZHU J, LI Y, YANG Y, LIU M. Anal. Bioanal. Chem., 2022, 414(20):6149- 6156.
17. [17]
  KATAOKA Y, FUJITA H, ENDOH T, SUGIMOTO N, KUWAHARA M. Molecules, 2020, 25(21):4936.KATAOKA Y, FUJITA H, ENDOH T, SUGIMOTO N, KUWAHARA M. Molecules, 2020, 25(21):4936.
18. [18]
  ZHOU Wei, LI Yun-Chao, FAN Lou-Zhen, LI Xiao-Hong. Acta. Phys-Chim. Sin., 2022, 38(4):91-98. 周蔚, 李运超, 范楼珍, 李晓宏. 物理化学学报, 2022, 38(4):91-98.
19. [19]
  LI Q S, PENG S Z, CHANG Y, YANG C, YANG M J, WANG D D, ZHOU X S, SHAO Y. J. Photochem. Photobio., A, 2022, 432:114071.LI Q S, PENG S Z, CHANG Y, YANG C, YANG M J, WANG D D, ZHOU X S, SHAO Y. J. Photochem. Photobio., A, 2022, 432:114071.
20. [20]
  ANDREEVA D V, TIKHOMIROV A S, SHCHEKOTIKHIN A E. Russ. Chem. Rev., 2021, 90(1):1-38.ANDREEVA D V, TIKHOMIROV A S, SHCHEKOTIKHIN A E. Russ. Chem. Rev., 2021, 90(1):1-38.
21. [21]
  CHANG Tian-Jun, WU Li, BIAN Wen-Xiu, BING Tao, ZHAO Tong-Qian, SHANGGUAN Di-Hua. Chin. J. Anal. Chem., 2020, 48(3):323-331. 常天俊, 武俐, 卞文秀, 邴涛, 赵同谦, 上官棣华. 分析化学, 2020, 48(3):323-331.
22. [22]
  VERMA S, RAVICHANDIRAN V, RANJAN N. Biochimie, 2021, 190:111-123.VERMA S, RAVICHANDIRAN V, RANJAN N. Biochimie, 2021, 190:111-123.
23. [23]
  LU S, WANG S, ZHAO J, SUN J, YANG X. Sci. China Chem., 2019, 63(3):404-410.LU S, WANG S, ZHAO J, SUN J, YANG X. Sci. China Chem., 2019, 63(3):404-410.
24. [24]
  SHEN F, QIAN H, CHENG Y, XIE Y, YU H, YAO W, PEI R, GUO Y, LI H W. Talanta, 2020, 206:120216.SHEN F, QIAN H, CHENG Y, XIE Y, YU H, YAO W, PEI R, GUO Y, LI H W. Talanta, 2020, 206:120216.
25. [25]
  ZHONG D, YANG K, WANG Y, YANG X. Talanta, 2019, 175:217-223.ZHONG D, YANG K, WANG Y, YANG X. Talanta, 2019, 175:217-223.
26. [26]
  HAN D, WEI C. Talanta, 2018, 181:24-31.HAN D, WEI C. Talanta, 2018, 181:24-31.
27. [27]
  GUO Y, XU P, HU H, ZHOU X, HU J. Talanta, 2013, 114:138-142.GUO Y, XU P, HU H, ZHOU X, HU J. Talanta, 2013, 114:138-142.

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沈阳化工大学材料科学与工程学院沈阳 110142

基于富G序列的无标记荧光传感器检测单链核酸

通讯作者: 陈青华,E-mail:2006007@zzuli.edu.cn; 姜利英,E-mail:jiangliying@zzuli.edu.cn

English

Label-free Fluorescent Sensor for Detection of Single-stranded Nucleic Acids Based on G-rich Sequence

Corresponding author: CHEN Qing-Hua, ; JIANG Li-Ying,

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

基于富G序列的无标记荧光传感器检测单链核酸

通讯作者: 陈青华,E-mail:2006007@zzuli.edu.cn; 姜利英,E-mail:jiangliying@zzuli.edu.cn

English

Label-free Fluorescent Sensor for Detection of Single-stranded Nucleic Acids Based on G-rich Sequence

Corresponding author: CHEN Qing-Hua, ; JIANG Li-Ying,

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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