Application of aptamer-based hybridization chain reaction in detections
-
摘要: 开发简单、灵敏的检测方法对医学诊断、环境监测、食品安全等领域的研究具有重要意义。适配体作为新型生物识别分子,与杂交链式反应(HCR)结合能实现高灵敏检测靶标物,形成一种新型检测模式。HCR是一种无需酶参与,常温下可自组装的扩增技术。基于适配体的HCR技术因具有高特异性、高灵敏性、操作简单、成本低廉等优点受到研究者们的广泛关注。本文主要介绍了HCR反应的基本原理及特征,重点综述了近年来基于适配体的HCR技术在蛋白质、酶活性、小分子及肿瘤细胞检测中的应用,探讨了该技术目前存在的主要问题,以期为建立高效率、高灵敏的适配体HCR检测系统提供理论参考。Abstract: Developing simple and sensitive detection methods is of great significance to researches on medical diagnosis, environmental analysis and food safety. As a new biological indicator molecule, aptamers are combined with hybridization chain reaction (HCR) to form a new detection model for sensitive detection of analytes. HCR is a typical amplification technology, which can be carried out without enzyme and self-assembled at room temperature. The aptamer-based HCR technology has be attracted great interest due to its high specificity and sensitivity, simple protocol and low cost. The study reviews basic features of HCR with an emphasis on the application of aptamer-based HCR in detection, such as proteins, enzyme activities, small molecules and tumor cells. Major existing problems of the technology are also discussed. The review is aimed to provide a theoretical reference for establishing a highly efficient and sensitive HCR detection system.
-
Key words:
- aptamer /
- hybridization chain reaction /
- high specificity /
- high sensitivity /
- detection
-
图 1 杂交链式反应的原理[7]
图 2 基于适配体HCR系统检测PDGF-BB的原理图[9]
图 3 扫描电化学显微镜检测示意图[17]
图 4 多分支HCR合成具有多个生物素标记和多个分支臂的长产物[34]
表 1 基于适配体的杂交链式反应用于检测蛋白质
检测物 信号单元 线性范围 检测限 参考文献 PDGF-BB SYBR Green I 5~5 000 pmol/L 1.25 pmol/L [10] AGR2 AuNPs 5~1 000 nmol/L 2.65 pmol/L [11] 糖类抗原CA125 AuNPs 0.10~10 U/mL 50 μU/mL [12] 外泌体 FAM 103~107 颗粒/mL 100颗粒/mL [13] 黏蛋白1 FAM 0.01~5 nmol/L 3.33 pmol/L [14] 黏蛋白16 MB 0.39~200 unit/ml 0.02 unit/ml [15] 血管内皮生长因子 H+ 0.80~480 pg/mL 0.50 pg/mL [16] 表 2 基于适配体的杂交链式反应用于检测酶活性
表 3 基于适配体的杂交链式反应用于检测小分子
检测物 信号单元 线性范围 检测限 参考文献 AFB1 鲁米诺/H2O2/HRP 0.50~40 ng/mL 0.20 ng/mL [22] AFB1 FAM 2~60 ng/mL 1.84 ng/mL [25] 赭曲霉毒素A TMB 0.01~0.32 nmol/L 0.01 nmol/L [26] 卡那霉素 MB 0.05~200 pmol/L 36 fmol/L [27] 卡那霉素 AuNPs 1.6~32 nmol/L 0.9 nmol/L [28] 多氯联苯 MB 1 × 10 – 5 ~10 ng/L 0.001ng/ml [29] ATP NMM 30~800 μmol/L 15 μmol/L [23] ATP GNP 10~500 nmol/L 10 nmol/L [30] Hg2 + [Ru(NH3)6]3 + 0.20~35 000 pmol/L 0.12 pmol/L [24] 腺苷 SYBR Green I 1~120 μmol/L 0.20 μmol/L [31] 8-羟基-2'-脱氧鸟苷 [Ru(NH3)6]3 + 100~10 000 fmol/L 24.34 fmol/L [32] -
[1] Gopinath SCB. Methods developed for SELEX[J]. Analytical and Bioanalytical Chemistry, 2007, 387(1): 171 – 182. [2] 李晓佩, 杨良嵘, 黄昆, 等. 核酸适配体在生化分离及检测领域中的研究进展[J]. 化工学报, 2013, 64(1): 233 – 242. doi: 10.3969/j.issn.0438-1157.2013.01.025 [3] Lorenz TC. Polymerase chain reaction: basic protocol plus troubleshooting and optimization strategies[J]. Journal of Visualized Experiments, 2012(63): 3998. [4] Zhu X, Xu HF, Zheng HY, et al. An ultrasensitive aptameric sensor for proteins based on hyperbranched rolling circle amplification[J]. Chemical Communications, 2013, 49(86): 10115 – 10117. doi: 10.1039/c3cc45521a [5] Li YB, Liu S, Zhao ZK, et al. Binding induced strand displacement amplification for homogeneous protein assay[J]. Talanta, 2017, 164: 196 – 200. doi: 10.1016/j.talanta.2016.11.047 [6] Bi S, Yue SZ, Zhang SS. Hybridization chain reaction: a versatile molecular tool for biosensing, bioimaging, and biomedicine[J]. Chemical Society Reviews, 2017, 46(14): 4281 – 4298. doi: 10.1039/C7CS00055C [7] Dirks RM, Pierce NA. Triggered amplification by hybridization chain reaction[J]. Proceedings of the National Academy of Sciences of the United States of America, 2004, 101(43): 15275 – 15278. doi: 10.1073/pnas.0407024101 [8] Venkataraman S, Dirks RM, Rothemund PWK, et al. An autonomous polymerization motor powered by DNA hybridiza-tion[J]. Nature Nanotechnology, 2007, 2(8): 490 – 494. doi: 10.1038/nnano.2007.225 [9] Song WQ, Zhu KL, Cao ZJ, et al. Hybridization chain reaction - based aptameric system for the highly selective and sensitive detection of protein[J]. Analyst, 2012, 137(6): 1396 – 1401. doi: 10.1039/c2an16232f [10] Wang XZ, Jiang AW, Hou T, et al. Enzyme - free and label - free fluorescence aptasensing strategy for highly sensitive detection of protein based on target - triggered hybridization chain reaction ampli-fication[J]. Biosensors and Bioelectronics, 2015, 70: 324 – 329. doi: 10.1016/j.bios.2015.03.053 [11] Li ZB, Miao XM, Cheng ZY, et al. Hybridization chain reaction coupled with the fluorescence quenching of gold nanoparticles for sensitive cancer protein detection[J]. Sensors and Actuators B:Chemical, 2017, 243: 731 – 737. doi: 10.1016/j.snb.2016.12.047 [12] Nie YT, Yang MY, Ding YL. Gold nanoparticle enhanced hybridization chain reaction as a method for signal amplification. Application to electrochemical immunodetection of the ovarian cancer biomarker carbohydrate antigen 125[J]. Microchimica Acta, 2018, 185(7): 331. doi: 10.1007/s00604-018-2869-4 [13] Shi LL, Ba L, Xiong Y, et al. A hybridization chain reaction based assay for fluorometric determination of exosomes using magnetic nanoparticles and both aptamers and antibody as recognition elements[J]. Microchimica Acta, 2019, 186(12): 796. doi: 10.1007/s00604-019-3823-9 [14] Ma C, Liu HY, Tian T, et al. A simple and rapid detection assay for peptides based on the specific recognition of aptamer and signal amplification of hybridization chain reaction[J]. Biosensors and Bioelectronics, 2016, 83: 15 – 18. doi: 10.1016/j.bios.2016.04.030 [15] Lu LS, Liu B, Leng JH, et al. Electrochemical mixed aptamer - antibody sandwich assay for mucin protein 16 detection through hybridization chain reaction amplification[J]. Analytical and Bioanalytical Chemistry, 2020, 412(26): 7169 – 7178. doi: 10.1007/s00216-020-02849-5 [16] Xu HF, Kou FX, Ye HZ, et al. Highly sensitive antibody - aptamer sensor for vascular endothelial growth factor based on hybridiza-tion chain reaction and pH meter/indicator[J]. Talanta, 2017, 175: 177 – 182. doi: 10.1016/j.talanta.2017.04.073 [17] Song WL, Xie XX, Sun WB, et al. Ultrasensitive electrochemical detection for thrombin using hybridization chain reaction with enzyme - amplification[J]. Analytica Chimica Acta, 2015, 860: 77 – 82. doi: 10.1016/j.aca.2014.12.029 [18] Jia LP, Zhao RN, Wang LJ, et al. Aptamer based electrochemical assay for protein kinase activity by coupling hybridization chain reaction[J]. Biosensors and Bioelectronics, 2018, 117: 690 – 695. doi: 10.1016/j.bios.2018.06.067 [19] Chen YX, Huang KJ, He LL, et al. Tetrahedral DNA probe coupling with hybridization chain reaction for competitive thrombin aptasensor[J]. Biosensors and Bioelectronics, 2018, 100: 274 – 281. doi: 10.1016/j.bios.2017.09.022 [20] Chang YY, Chai YQ, Xie SB, et al. Cleavage - based hybridization chain reaction for electrochemical detection of thrombin[J]. Analyst, 2014, 139(17): 4264 – 4269. doi: 10.1039/C4AN00712C [21] 曹亚, 康明扬, 陈红, 等. 基于杂交链式反应辅助多重信号放大的端粒酶灵敏检测[J]. 分析化学, 2017, 45(12): 1903 – 1908. doi: 10.11895/j.issn.0253-3820.171340 [22] Yao YY, Wang HX, Wang XZ, et al. Development of a chemiluminescent aptasensor for ultrasensitive and selective detection of aflatoxin B1 in peanut and milk[J]. Talanta, 2019, 201: 52 – 57. doi: 10.1016/j.talanta.2019.03.109 [23] Chen QG, Guo QQ, Chen Y, et al. An enzyme - free and label - free fluorescent biosensor for small molecules by G - quadruplex based hybridization chain reaction[J]. Talanta, 2015, 138: 15 – 19. doi: 10.1016/j.talanta.2015.02.002 [24] Bao T, Wen W, Zhang XH, et al. An exonuclease-assisted amplification electrochemical aptasensor for Hg2 + detection based on hybridization chain reaction[J]. Biosensors and Bioelectronics, 2015, 70: 318 – 323. doi: 10.1016/j.bios.2015.03.065 [25] 黄玉坤, 陶璇, 邵坤, 等. 基于适配体杂交链式反应检测郫县豆瓣中黄曲霉毒素B1[J]. 食品科学, 2020, 41(22): 301 – 307. doi: 10.7506/spkx1002-6630-20190903-037 [26] Wang CK, Dong XY, Liu Q, et al. Label-free colorimetric aptasensor for sensitive detection of ochratoxin A utilizing hybridization chain reaction[J]. Analytica Chimica Acta, 2015, 860: 83 – 88. doi: 10.1016/j.aca.2014.12.031 [27] Zeng RJ, Su LS, Luo ZB, et al. Ultrasensitive and label-free electrochemical aptasensor of kanamycin coupling with hybridi-zation chain reaction and strand-displacement amplification[J]. Analytica Chimica Acta, 2018, 1038: 21 – 28. doi: 10.1016/j.aca.2018.07.010 [28] 田润, 陶晴, 卞晓军, 等. 基于杂交链式反应的适配体传感器用于卡那霉素的比色检测[J]. 分析化学, 2020, 48(5): 608 – 614. [29] Han T, Wang SZ, Sheng FF, et al. Target triggered ultrasensitive electrochemical polychlorinated biphenyl aptasensor based on DNA microcapsules and nonlinear hybridization chain reaction[J]. Analyst, 2020, 145(10): 3598 – 3604. doi: 10.1039/D0AN00065E [30] Li S, Shang XX, Liu J, et al. A universal colorimetry for nucleic acids and aptamer-specific ligands detection based on DNA hybridization amplification[J]. Analytical Biochemistry, 2017, 528: 47 – 52. doi: 10.1016/j.ab.2017.04.013 [31] Feng CJ, Hou Z, Jiang W, et al. Binding induced colocalization activated hybridization chain reaction on the surface of magnetic nanobead for sensitive detection of adenosine[J]. Biosensors and Bioelectronics, 2016, 86: 966 – 970. doi: 10.1016/j.bios.2016.07.108 [32] Jia LP, Feng Z, Zhao RN, et al. Enzyme-free and triple-amplified electrochemical sensing of 8-hydroxy-2'-deoxyguanosine by three kinds of short pDNA-driven catalyzed hairpin assemblies followed by a hybridization chain reaction[J]. Analyst, 2020, 145(10): 3605 – 3611. doi: 10.1039/D0AN00233J [33] Zhang Y, Chen ZW, Tao Y, et al. Hybridization chain reaction engineered dsDNA for Cu metallization: an enzyme-free platform for amplified detection of cancer cells and microRNAs[J]. Chemical Communications, 2015, 51(57): 11496 – 11499. doi: 10.1039/C5CC03144C [34] Zhou GB, Lin MH, Song P, et al. Multivalent capture and detection of cancer cells with DNA nanostructured biosensors and multibranched hybridization chain reaction amplification[J]. Analytical Chemistry, 2014, 86(15): 7843 – 7848. doi: 10.1021/ac502276w [35] Yuan BY, Guo LY, Yin K, et al. Highly sensitive and specific detection of tumor cells based on a split aptamer-triggered dual hybridization chain reaction[J]. Analyst, 2020, 145(7): 2676 – 2681. doi: 10.1039/C9AN02476J [36] Li L, Jiang HS, Meng XX, et al. Highly sensitive detection of cancer cells via split aptamer mediated proximity-induced hybridization chain reaction[J]. Talanta, 2021, 223: 121724. doi: 10.1016/j.talanta.2020.121724 [37] Tang JL, Lei YL, He XX, et al. Recognition-driven remodeling of dual-split aptamer triggering in situ hybridization chain reaction for activatable and autonomous identification of cancer cells[J]. Analytical Chemistry, 2020, 92(15): 10839 – 10846. doi: 10.1021/acs.analchem.0c02524