Advertisement

Analytical and Bioanalytical Chemistry

, Volume 411, Issue 1, pp 139–146 | Cite as

An aptamer biosensor for leukemia marker mRNA detection based on polymerase-assisted signal amplification and aggregation of illuminator

  • Meng Zhang
  • Fenyue Zhou
  • Deqi Zhou
  • Dongli Chen
  • Hong Hai
  • Jianping Li
Paper in Forefront

Abstract

A novel electrochemical luminescence (ECL) aptamer biosensor via polymerase amplification is constructed for label-free detection of leukemia marker mRNA (miR-16). In order to achieve the ultrasensitive detection of the target mRNA, the cyclic target chain displacement polymerization of leukemia marker mRNA assisted with Klenow fragment of DNA polymerase is employed. The determination is carried out by recording the ECL emission of pyridine ruthenium (Ru(bpy)32+) complexes embedded into the assistance DNA (ADNA) loaded on the nanogold surface, after the hybridization reaction between the probe DNA (PDNA) and the remaining sequence of the CP’s stem part, and the formation of a core-shell sun-like structure. The mercapto-modified capture DNA (CP) is immobilized on the surface of a magneto-controlled glassy carbon electrode by Au-S bond. The CP is opened and hybridized with the target mRNA to form double-stranded DNA. In the presence of polymerase, primer DNA, and bases (dNTPs), the primer chain gets access to its complementary sequence of the stem part and then triggers a polymerization of the DNA strand, leading to the release of mRNA and starting the next polymerization cycle. Finally, the composite of PDNA-covered and ADNA-covered (embedded with Ru(bpy)32+) gold nanoparticles (hereafter called AuNPs@(PDNA+ADNA-Ru(bpy)32+) is added, and the ECL intensity is recorded. Because of the polymerization cycle and the aggregation of the illuminator of Ru(bpy)32+, the detected signal is amplified significantly. The results showed that the corresponding ECL signal has a good linear relationship with a logarithm of target mRNA concentration in the range of 1 × 10−16 to 1 × 10−7 mol/L, with a detection limit of 4.3 × 10−17 mol/L. The mRNA spiked in the human serum sample is determined, and the recoveries are from 97.2 to 102.0%. This sensor demonstrates good selectivity, stability, and reproducibility.

Graphical abstract

Keywords

Aptamer sensor Polymerase Electrochemiluminescence Label-free Cyclic amplification Aggregation effect 

Notes

Funding information

This research is financially supported by the National Natural Science Foundation of China (No.21765006), the Guangxi Natural Science Foundation, China (No. 2015GXNSFFA139005), and the High-Level Innovation Teams of Guangxi Colleges and Universities and Outstanding Scholars Program (Guijiaoren[2014]49).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

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.

Supplementary material

216_2018_1424_MOESM1_ESM.pdf (241 kb)
ESM 1 (PDF 204 kb)

References

  1. 1.
    Musto P, Negrini M, De LL, Cuneo A. Dissecting chronic lymphocytic leukemia with 13q-using microRNA expression profile: editorial for the paper “miRNA expression profile of chronic lymphocytic leukemia patients with 13q deletion”. Leuk Res. 2016;47:114–5.CrossRefGoogle Scholar
  2. 2.
    Achille NJ, Othus M, Phelam K, Zhang SB, Cooper K, Godwin JE, et al. Association between early promoter-specific DNA methylation changes and outcome in older acute myeloid leukemia patients. Leuk Res. 2016;42:68–74.CrossRefGoogle Scholar
  3. 3.
    Buckley SA, Jimenez SD, Othus M, Walter RB, Lee SJ. Quality of life from the perspective of the patient with acute myeloid leukemia. Cancer. 2018;124:145–52.CrossRefGoogle Scholar
  4. 4.
    Setiadi A, Owen D, Tsang A, Milner R, Vercautern S. The significance of peripheral blood minimal residual disease to predict early disease response in patients with B–cell acute lymphoblastic leukemia. Int J Lab Hematol. 2016;38:527–34.CrossRefGoogle Scholar
  5. 5.
    Boyerinas B, Zafrir M, Yesilkanal AE, Price TT, Hyjek EM, Sipkins DA. Adhesion to osteopontin in the bone marrow niche regulates lymphoblastic leukemia cell dormancy. Blood. 2013;121:4821–31.CrossRefGoogle Scholar
  6. 6.
    Jenderny J, Goldmann C, Thede R, Ebrecht M, Korioth F. Detection of clonal aberrations by cytogenetic analysis after different culture methods and by FISH in 129 patients with chronic lymphocytic leukemia. Cytogenet Genome Res. 2014;144:163–8.CrossRefGoogle Scholar
  7. 7.
    Esfandyarpour R, Yang L, Koochak Z, Harris JS, Davis RW. Nanoelectronic three-dimensional (3D) nanotip sensing array for real-time, sensitive, label-free sequence specific detection of nucleic acids. Biomed Microdevices. 2016;18:1–10.CrossRefGoogle Scholar
  8. 8.
    Kitamura M, Aragane M, Nakamura K, Watanabe K, Sasaki Y. Development of loop-mediated isothermal amplification (LAMP) assay for rapid detection of Cannabis sativa. Biol Pharm Bull. 2016;39:1144–9.CrossRefGoogle Scholar
  9. 9.
    Zhang M, Hai H, Zhou FY, Zhong JC, Li JP. Electrochemical luminescent DNA sensor based on polymerase-assisted signal amplification. Chin J Anal Chem. 2018;46:203–9.CrossRefGoogle Scholar
  10. 10.
    Martens ES, Böttcher R, Croce CM, Jenster G, Visakorpi T, Calin GA. Long noncoding RNA in prostate, bladder, and kidney cancer. Eur Urol. 2014;65:1140–51.CrossRefGoogle Scholar
  11. 11.
    Yang LM, Li J, Pan W, Wang HY, Li N, Tang BY. Fluorescence and photoacoustic dual-mode imaging of tumor-related mRNA with a covalent linkage-based DNA nanoprobe. Chem Commun. 2018;54:3656–9.CrossRefGoogle Scholar
  12. 12.
    Miao P, Jiang YT, Zhang T, Huang Y, Yang YG. Electrochemical sensing of attomolar miRNA combining cascade strand displacement polymerization and reductant-mediated amplification. Chem Commun. 2018;54:7366–9.CrossRefGoogle Scholar
  13. 13.
    Calin GA, Dumitru CD, Shimizu M, Bichi R, Zupo S, Noch E, et al. Frequent deletions and down-regulation of micro-RNA genes miR15 and miR16 at 13q14 in chronic lymphocytic leukemia. Proc Natl Acad Sci. 2002;99:15524–9.CrossRefGoogle Scholar
  14. 14.
    Enfield KSS, Martinez VD, Marshall EA, Stewart GL, Kung SH, Enterina JR, et al. Deregulation of small non-coding RNAs at the DLK1-DIO3 imprinted locus predicts lung cancer patient outcome. Oncotarget. 2016;7:80957–66.CrossRefGoogle Scholar
  15. 15.
    Yu MM, Yao S, Luo KM, Mu QF, Yu Y, Luo GH, et al. Apolipoprotein M increases the expression of vitamin D receptor mRNA in colorectal cancer cells detected with duplex fluorescence reverse transcription-quantitative polymerase chain reaction. Mol Med Rep. 2017;16:1167–72.CrossRefGoogle Scholar
  16. 16.
    Sanyal S, Siriwardena AK, Byers R. Measurement of indicator genes using global complementary DNA (cDNA) amplification, by polyadenylic acid reverse transcriptase polymerase chain reaction (poly A RT-PCR): a feasibility study using paired samples from tissue and ductal juice in patients undergoing pancreatoduodenectomy. Pancreatology. 2018;18:458–62.CrossRefGoogle Scholar
  17. 17.
    Lin X, Chen Y. Identification of potentially functional circRNA-miRNA-mRNA regulatory network in hepatocellular carcinoma by integrated microarray analysis. Med Sci Monit. 2018;24:70–8.Google Scholar
  18. 18.
    Ma Y, Shi L, Zheng C. Microarray analysis of lncRNA and mRNA expression profiles in mice with allergic rhinitis. Int J Pediatr Otorhinolaryngol. 2018;104:58–65.CrossRefGoogle Scholar
  19. 19.
    Wang S, Zhang LQ, Wan S, Gansiz S, Cui C, Liu Y, et al. Aptasensor with expanded nucleotide using DNA nanotetrahedra for electrochemical detection of cancerous exosomes. ACS Nano. 2017;11:3943–9.CrossRefGoogle Scholar
  20. 20.
    Li JP, Sun M, Wei XP, Zhang LM, Zhang Y. An electrochemical aptamer biosensor based on “gate-controlled” effect using β-cyclodextrin for ultra-sensitive detection of trace mercury. Biosens Bioelectron. 2015;74:423–6.CrossRefGoogle Scholar
  21. 21.
    Zhou FY, Hai H, Yuan YL, Li JP. Ultrasensitive electrochemiluminescence biosensor for mRNA based on polymerase assisted signal amplification. Electroanalysis. 2017;29:983–9.CrossRefGoogle Scholar
  22. 22.
    Qin GX, Zhao SL, Huang Y, Jiang J, Liu YM. A sensitive gold nanoparticles sensing platform based on resonance energy transfer for chemiluminescence light on detection of biomolecules. Biosens Bioelectron. 2013;46:119–23.CrossRefGoogle Scholar
  23. 23.
    Wang LP, Huang YB, Lai YH. Surface enhanced Raman scattering activity of dual-functional Fe3O4/Au composites. Appl Surf Sci. 2018;435:290–6.CrossRefGoogle Scholar
  24. 24.
    Yao Y, Xue M, Zhang ZB, Zhang MM, Wang Y, Huang FH. Gold nanoparticles stabilized by an amphiphilic pillar [5] arene: preparation, self-assembly into composite microtubes in water and application in green catalysis. Chem Sci. 2013;4:3667–72.CrossRefGoogle Scholar
  25. 25.
    Ding QW, Zhan QQ, Zhou XM, Zhang T, Xing D. Theranostic upconversion nanobeacons for tumor mRNA ratiometric fluorescence detection and imaging-monitored drug delivery. Small. 2016;12:5944–53.CrossRefGoogle Scholar
  26. 26.
    Ali MM, Li F, Zhang ZQ, Zhang KX, Kang DK, Ankrum JA, et al. Rolling circle amplification: a versatile tool for chemical biology, materials science and medicine. Chem Soc Rev. 2014;43:3324–41.CrossRefGoogle Scholar
  27. 27.
    Alsage OA, Kumar S, Zhu BC, Sejdic JT, McNatty KP, Hodgkiss JM. Ultrasensitive colorimetric detection of 17β-estradiol: the effect of shortening DNA aptamer sequences. Anal Chem. 2015;87:4201–9.CrossRefGoogle Scholar
  28. 28.
    Zhao FL, Xie QY, Xu MF, Wang SY, Zhou JY, Liu F. RNA aptamer based electrochemical biosensor for sensitive and selective detection of cAMP. Biosens Bioelectron. 2015;66:238–43.CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Guangxi Key Laboratory of Electrochemical and Magnetochemical Functional Materials, College of Chemistry and BioengineeringGuilin University of TechnologyGuilinChina
  2. 2.College of Biological SciencesUniversity of California – DavisDavisUSA

Personalised recommendations