Skip to main content
SpringerLink
Log in

A three-line lateral flow biosensor for logic detection of microRNA based on Y-shaped junction DNA and target recycling amplification

  • Research Paper
  • Published:
Analytical and Bioanalytical Chemistry Aims and scope Submit manuscript

Abstract

A rapid, sensitive, and accurate detection strategy for microRNA 16 (miR-16) was developed, which combined the convenience of lateral flow biosensors (LFBs), the design flexibility of Y-shaped junction DNA probe, and the enhancement ability of endonuclease-assisted target recycling amplification. The system is composed of a molecular beacon (MB) probe, an assistant probe, and endonuclease Nt.BbvCI, which plays the role of signal translation and amplification. In the presence of the target microRNAs (miRNAs), three chains of nucleic acid could hybridize with each other to form a Y-shaped junction structure, which could be recognized by the endonuclease Nt.BbvCI. The MB probe was efficiently cleaved by endonuclease and produced two new DNA fragments, while the regenerated assistant probe and target were hybridized to another MB probe and entered into the next cycle of the amplification. In this way, the detection of the readily biodegradable miRNA was turned into the detection of two DNA fragments in the LFB. Meanwhile, the detection of two different DNAs would improve the accuracy and effectively avoid false results. The amplified products containing DNA fragments were then applied to the lateral flow nucleic acid biosensor (LFNAB) with two test zones, on which specific DNA probes were designed. The formed DNA-DNA/gold nanoparticle (GNP) conjugates were captured and accumulated to produce two red bands in two test zones. The logic judgment of the two test zones provided more accurate and convincing results. Under optimal conditions, the visual detection limit of miR-16 in aqueous solutions was 0.1 pM, which is 100–1000 times lower than that of visual or colorimetric methods in the literature. It could be used for on-field and point-of-care testing and meet the urgent demand of sensitive and selective miRNA detection in remote rural areas without costly equipment. The system displayed good universality, compatibility, high specificity, and stability of miRNA detection, which revealed significant potentiality in biomedical diagnosis.

A simple and rapid approach for microRNA-16 (miR-16) detection was developed based on Y-shaped junction DNA probe and the endonuclease-assisted target recycling amplification using lateral flow biosensors.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Krol J, Loedige I, Filipowicz W. The widespread regulation of microRNA biogenesis, function and decay. Nat Rev Genet. 2010;11:597–610.

    CAS  Google Scholar 

  2. Lu J, Getz G, Miska EA, Alvarez-Saavedra E, Lamb J, Peck D, et al. MicroRNA expression profiles classify human cancers. Nature. 2005;435:834–8.

    Article  CAS  Google Scholar 

  3. Taganov KD, Boldin MP, Chang KJ, Baltimore D. NF-kB-dependent induction of microRNA miR-146, an inhibitor targeted to signaling proteins of innate immune responses. Proc Natl Acad Sci. 2006;103:12481–6.

    Article  CAS  Google Scholar 

  4. Duan R, Zuo X, Wang S, Quan X, Chen D, Chen Z, et al. Lab in a tube: ultrasensitive detection of microRNAs at the single-cell level and in breast cancer patients using quadratic isothermal amplification. J Am Chem Soc. 2013;135:4604–7.

    Article  CAS  Google Scholar 

  5. Dong H, Lei J, Ding L, Wen Y, Ju H, Zhang X. MicroRNA: function, detection, and bioanalysis. Chem Rev. 2013;113:6207–33.

    Article  CAS  Google Scholar 

  6. Liu G, Lin YY, Wang J, Wu H, Wai CM, Lin Y. Disposable electrochemical immunosensor diagnosis device based on nanoparticle probe and immunochromatographic strip. Anal Chem. 2007;79:7644–53.

    Article  CAS  Google Scholar 

  7. Parolo C, Merkoci A. Paper-based nanobiosensors for diagnostics. Chem Soc Rev. 2013;42:450–7.

    Article  CAS  Google Scholar 

  8. Xu H, Chen J, Birrenkott J, Zhao JX, Takalkar S, Baryeh K, et al. Gold-nanoparticle-decorated silica nanorods for sensitive visual detection of proteins. Anal Chem. 2014;86:7351–9.

    Article  CAS  Google Scholar 

  9. Oh YK, Joung HA, Han HS, Suk HJ, Kim MG. A three-line lateral flow assay strip for the measurement of C-reactive protein covering a broad physiological concentration range in human sera. Biosens Bioelectron. 2014;61:285–9.

    Article  CAS  Google Scholar 

  10. Zhu X, Shah P, Stoff S, Liu H, Li CZ. A paper electrode integrated lateral flow immunosensor for quantitative analysis of oxidative stress induced DNA damage. Analyst. 2014;139:2850–7.

    Article  CAS  Google Scholar 

  11. Dong H, Hao K, Tian Y, Jin S, Lu H, Zhou SF, et al. Label-free and ultrasensitive microRNA detection based on novel molecular beacon binding readout and target recycling amplification. Biosens Bioelectron. 2014;53:377–83.

    Article  CAS  Google Scholar 

  12. Gao X, Xu H, Baloda M, Gurung AS, Xu L-P, Wang T, et al. Visual detection of microRNA with lateral flow nucleic acid biosensor. Biosens Bioelectron. 2014;54:578–84.

    Article  CAS  Google Scholar 

  13. Dong H, Meng X, Dai W, Cao Y, Lu H, Zhou S, et al. Highly sensitive and selective microRNA detection based on DNA-bio-bar-code and enzyme-assisted strand cycle exponential signal amplification. Anal Chem. 2015;87:4334–40.

    Article  CAS  Google Scholar 

  14. Gao X, Xu LP, Wu T, Wen Y, Ma X, Zhang X. An enzyme-amplified lateral flow strip biosensor for visual detection of microRNA-224. Talanta. 2016;146:648–54.

    Article  CAS  Google Scholar 

  15. Liu M, Hui C, Zhang Q, Gu J, Kannan B, Jahanshahi-anhubi S, et al. Target-induced and equipment-free DNA amplification with a simple paper device. Angew Chem Int Ed. 2016;128:2759–63.

    Article  Google Scholar 

  16. Kor K, Turner A, Zarei K, Atabati M, Beni V, Mak W. Structurally responsive oligonucleotide-based single-probe lateral-flow test for detection of miRNA-21 mimics. Anal Bioanal Chem. 2016;408:1475–85.

    Article  CAS  Google Scholar 

  17. Takalkar S, Xu H, Chen J, Baryeh K, Qiu W, Zhao J, et al. Gold nanoparticle coated silica nanorods for sensitive visual detection of microRNA on a lateral flow strip biosensor. Anal Sci. 2016;32:617–21.

    Article  CAS  Google Scholar 

  18. Mao X, Ma Y, Zhang A, Zhang L, Zeng L, Liu G. Disposable nucleic acid biosensors based on gold nanoparticle probes and lateral flow strip. Anal Chem. 2009;81:1660–8.

    Article  CAS  Google Scholar 

  19. He Y, Zeng K, Gurung AS, Baloda M, Xu H, Zhang X, et al. Visual detection of single-nucleotide polymorphism with hairpin oligonucleotide-functionalized gold nanoparticles. Anal Chem. 2010;82:7169–77.

    Article  CAS  Google Scholar 

  20. Yang D, Hartman MR, Derrien TL, Hamada S, An D, Yancey KG, et al. DNA materials: bridging nanotechnology and biotechnology. Acc Chem Res. 2014;47:1902–11.

    Article  CAS  Google Scholar 

  21. Chen J, Zhou S, Wen J. Concatenated logic circuits based on a three-way DNA junction: a keypad-lock security system with visible readout and an automatic reset function. Angew Chem Int Ed. 2015;127:456–60.

    Article  Google Scholar 

  22. Liu S, Gong H, Sun X, Liu T, Wang L. A programmable Y-shaped junction scaffold-mediated modular and cascade amplification strategy for the one-step, isothermal and ultrasensitive detection of target DNA. Chem Commun. 2015;51:17756–9.

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by the NSFC (21171019, 51373023, 21073203) and the Fundamental Research Funds for the Central Universities.

Author information

Authors and Affiliations

  1. Research Center for Bioengineering and Sensing Technology, University of Science and Technology Beijing, Beijing, 100083, China

    Yan Huang, Wenqian Wang, Tingting Wu, Li-Ping Xu, Yongqiang Wen & Xueji Zhang

Authors
  1. Yan Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Wenqian Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Tingting Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Li-Ping Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yongqiang Wen
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Xueji Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Yongqiang Wen or Xueji Zhang.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Electronic supplementary material

Below is the link to the electronic supplementary material.

ESM 1

(PDF 245 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Huang, Y., Wang, W., Wu, T. et al. A three-line lateral flow biosensor for logic detection of microRNA based on Y-shaped junction DNA and target recycling amplification. Anal Bioanal Chem 408, 8195–8202 (2016). https://doi.org/10.1007/s00216-016-9925-x

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00216-016-9925-x

Keywords

Search

Navigation