Microchimica Acta

, 186:628 | Cite as

Ultrasensitive electroluminescence biosensor for a breast cancer marker microRNA based on target cyclic regeneration and multi-labeled magnetized nanoparticles

  • Dongli Chen
  • Meng Zhang
  • Fenyue Zhou
  • Hong HaiEmail author
  • Jianping LiEmail author
Original Paper


An electrochemiluminescent (ECL) biosensor is described for the determination of the breast cancer biomarker microRNA. The method is based on the amplification via target cyclic regeneration through a system of hairpin DNA probes, primers, and Klenow fragment of DNA polymerases combined with CdTe quantum dots (QDs) and gold nanoparticles. The assay is performed by exploiting the luminescence properties of CdTe-QDs and K2S2O8 as a co-reactive agent to increase the ECL signal. It was successfully applied to ECL-based detection of a 20-mer microRNA. The sensor has a linear response in the 0.1 fM to 0.2 pM microRNA concentration range and a detection limit as low as 33 aM. The assay has been applied to the determination of microRNA spiked in serum samples, and recoveries ranged from 94.4 to 100.5%.

Graphical abstract

A novel electroluminescence biosensor based on the amplification of target cyclic regeneration is described. It is achieved by using a system of hairpin DNA probes, primers, and Klenow fragment of DNA polymerases combined with CdTe QDs and Au NPs, and was successfully applied to microRNA detection.


Biosensor Electrochemiluminescence Breast cancer MicroRNA Quantum dots Polymerase Enzyme-assistant Signal amplification Cyclic regeneration Serum samples 



The authors gratefully acknowledge the financial support from National Natural Science Foundation of China (No. 21765006), the Natural Science Foundation of Guangxi Province of China (2015GXNSFFA139005; 2018GXNSFAA050023), Guangxi Colleges and Universities Key Laboratory of Food Safety and Detection, High Level Innovation Teams of Guangxi Colleges & Universities and Outstanding Scholars Program (Guijiaoren[2014]49) for this research project.

Compliance with ethical standards

The author(s) declare that they have no competing interests.

Ethical standards and informed consent

The study and experimental sections were approved by the Hospital of Guilin University of Technology. Human serum samples used in this study do not have any identifying information about all the participants that provided written informed consent.


  1. 1.
    Teengam P, Siangproh W, Tuantranont A, Vilaivan T, Chailapakul O, Henry CS (2018) Electrochemical impedance-based DNA sensor using pyrrolidinyl peptide nucleic acids for tuberculosis detection. Anal Chim Acta 1044:102–109CrossRefGoogle Scholar
  2. 2.
    Schulz D, Zanotelli VRT, Fischer JR, Schapiro D, Engler S, Lun XK, Jackson HW, Bodenmiller (2018) Simultaneous multiplexed imaging of mRNA and proteins with subcellular resolution in breast Cancer tissue samples by mass cytometry. Cell Syst 6:25–36CrossRefGoogle Scholar
  3. 3.
    Hirschberger K, Jarzebinska A, Kessel E, Kretzschmann V, Aneja MK, Dohmen C, Herrmann-Janson A, Wagner E, Plank C, Rudolph C (2018) Exploring cytotoxic mRNAs as a novel class of anti-cancer biotherapeutics. Mol Ther-Meth Clin D 8:141–151CrossRefGoogle Scholar
  4. 4.
    Zhao B, Zhao Y, Sun Y, Niu HT, Sheng L, Huang DF, Li L (2018) Alterations in mRNA profiles of trastuzumab-resistant Her-2-positive breast cancer. Mol Med Rep 18:139–146PubMedPubMedCentralGoogle Scholar
  5. 5.
    Antonio MD, Tamayo P, Mesirov JP, Frazer KA (2016) Kataegis expression signature in breast cancer is associated with late onset, better prognosis, and higher HER2 levels. Cell Rep 16:672–683CrossRefGoogle Scholar
  6. 6.
    Imani S, Zhang X, Hosseinifard H, Fu SY, Fu JJ (2017) The diagnostic role of microRNA-34a in breast cancer: a systematic review and meta-analysis. Oncotarget 8:23177–23187PubMedPubMedCentralGoogle Scholar
  7. 7.
    Zhang HY, Liang F, Zhang JW, Wang F, Wang L, Kang XG (2017) Effects of long noncoding RNA-ROR on tamoxifen resistance of breast cancer cells by regulating microRNA-205. Cancer Chemother Pharmacol 79:327–337CrossRefGoogle Scholar
  8. 8.
    Vilela P, El–Sagheer A, Millar TM, Brown T, Muskens OL, Kanaras AG (2017) Graphene oxide-Upconversion nanoparticle based optical sensors for targeted detection of mRNA biomarkers present in Alzheimer’s disease and prostate Cancer. ACS Sens 2:52–56CrossRefGoogle Scholar
  9. 9.
    Trayhurn P, Duncan JS, Nestor A, Thomas MEA, Eastmond NC, Vernon RD (2010) Rapid chemiluminescent detection of mRNAs on northern blots with digoxigenin end-labelled oligonucleotides. Electrophoresis 16:341–344CrossRefGoogle Scholar
  10. 10.
    Lee JM, Cho H, Jung Y (2016) Fabrication of a structure-specific RNA binder for Array detection of label-free MicroRNA. Angew Chem Int Edit 49:8662–8665CrossRefGoogle Scholar
  11. 11.
    Zhang M, Zhou FY, Zhou DQ, Chen DL, Hai H, Li JP (2019) An aptamer biosensor for leukemia marker mRNA detection based on polymerase-assisted signal amplification and aggregation of illuminator. Anal Bioanal Chem 411:139–146CrossRefGoogle Scholar
  12. 12.
    Cassol D, Cruz FP, Espindola K, Espindola K, Mangeon A, Muller C, Loureiro ME, Correa RL, Sachetto-Martins G (2016) Identification of reference genes for quantitative RT-PCR analysis of microRNAs and mRNAs in castor bean (Ricinus communis L.) under drought stress. Plant Physiol Biochem 106:101–107CrossRefGoogle Scholar
  13. 13.
    He L, Lu DQ, Liang H, Xie ST, Luo C, Hu MM, Xu LJ, Zhang XB, Tan WH (2017) Fluorescence resonance energy transfer-based DNA tetrahedron nanotweezer for highly reliable detection of tumor-related mRNA in living cells. ACS Nano 11:4060–4066CrossRefGoogle Scholar
  14. 14.
    Luan M, Li N, Pan W, Yang L, Yu Z, Tang B (2016) Simultaneous detection of multiple targets involved in the PI3K/AKT pathway for investigating cellular migration and invasion with a multicolor fluorescent nanoprobe. Chem Commun 53:356–359CrossRefGoogle Scholar
  15. 15.
    Liu RJ, Zhao JJ, Huang ZR, Zhang LL, Shi BF, Zou MB, Zhao SL (2017) Nitrogen and phosphorus co-doped graphene quantum dots as a nano-sensor for highly sensitive and selective imaging detection of nitrite in live cell. Sensors Actuators B Chem 240:604–612CrossRefGoogle Scholar
  16. 16.
    Ban F, Shi H, Feng C, Mao X, Yin Y, Zhu X (2016) A one-pot strategy for the detection of proteins based on sterically and allosterically tunable hybridization chain reaction. Biosens Bioelectron 86:219–224CrossRefGoogle Scholar
  17. 17.
    Yu X, Zhang ZL, Zheng SY (2015) Highly sensitive DNA detection using cascade amplification strategy based on hybridization chain reaction and enzyme-induced metallization. Biosens Bioelectron 66:520–526CrossRefGoogle Scholar
  18. 18.
    Zhang KY, Lv SZ, Lin ZZ, Li MJ, Tang DP (2018) Bio-bar-code-based photoelectrochemical immunoassay for sensitive detection of prostate-specific antigen using rolling circle amplification and enzymatic biocatalytic precipitation. Biosens Bioelectron 101:159–166CrossRefGoogle Scholar
  19. 19.
    Shen QM, Fan MX, Yang Y, Zhang H (2016) Electrochemical DNA sensor–based strategy for sensitive detection of DNA demethylation and DNA demethylase activity. Anal Chim Acta 934:66–71CrossRefGoogle Scholar
  20. 20.
    Huang KJ, Shuai HL, Zhang JZ (2016) Ultrasensitive sensing platform for platelet-derived growth factor BB detection based on layered molybdenum selenide–graphene composites and ExonucleaseIII assisted signal amplification. Biosens Bioelectron 77:69–75CrossRefGoogle Scholar
  21. 21.
    Gan XR, Zhao HM, Chen S, Quan X (2015) Electrochemical DNA sensor for specific detection of picomolar hg (II) based on exonuclease III-assisted recycling signal amplification. Analyst 140:2029–2036CrossRefGoogle Scholar
  22. 22.
    Zhou FY, Hai H, Yuan YL, Li JP (2017) Ultrasensitive Electrochemiluminescence biosensor for mRNA based on polymerase assisted signal amplification. Electroanal 29:983–989CrossRefGoogle Scholar
  23. 23.
    Wang LP, Huang YB, Lai H (2018) Surface enhanced Raman scattering activity of dual-functional Fe3O4/Au composites. Appl Surf Sci 435:290–296CrossRefGoogle Scholar
  24. 24.
    Xue M, Wang W (2016) Facile one-pot synthesis of water-soluble β-cyclodextrin coated CdTe quantum dots. Mater Lett 166:97–100CrossRefGoogle Scholar
  25. 25.
    Murayama T, Haruta M (2016) Preparation of gold nanoparticles supported on Nb2O5 by deposition precipitation and deposition reduction methods and their catalytic activity for CO oxidation. Chin J Catal 37:1694–1701CrossRefGoogle Scholar
  26. 26.
    Jackson SR, Wong AC, Travis AR, Catrina IE, Bratu DP, Wright DW, Jayagopal A (2016) Chapter four–applications of hairpin DNA-functionalized gold nanoparticles for imaging mRNA in living cells. Methods Enzymol 572:87–103CrossRefGoogle Scholar
  27. 27.
    Zhang XF, Xu HM, Han L, Li NB, Luo HQ (2018) A Thioflavin T-induced G-Quadruplex fluorescent biosensor for target DNA detection. Anal Sci 34:149–153CrossRefGoogle Scholar
  28. 28.
    Lee CY, Fan HT, Hsieh YZ (2018) Disposable aptasensor combining functional magnetic nanoparticles with rolling circle amplification for the detection of prostate-specific antigen. Sensors Actuators B Chem 255:341–347CrossRefGoogle Scholar
  29. 29.
    Lei Y, Wang K, Wu SY, Huang DD, Dai M, Zheng YJ, Sun ZL, Chen YZ, Lin XH, Liu AL (2018) 2’-Fluoro ribonucleic acid modified DNA dual-probe sensing strategy for enzyme-amplified electrochemical detection of double-strand DNA of PML/RARα related fusion gene. Biosens Bioelectron 112:170–176CrossRefGoogle Scholar
  30. 30.
    Ma RN, Wang LL, Wang HF, Jia LP, Zhang W, Shang L, Xue QW, Jia WL, Liu QY, Wang HS (2018) Highly sensitive ratiometric electrochemical DNA biosensor based on homogeneous exonuclease III-assisted target recycling amplification and one-step triggered dual-signal output. Sensors Actuators B Chem 269:173–179CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2019

Authors and Affiliations

  1. 1.College of Chemistry and BioengineeringGuilin University of TechnologyGuangxiChina
  2. 2.Guangxi Key Laboratory of Electrochemical and Magnetochemical Function MaterialsGuangxiChina

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