Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

Plasmonic ELISA for the ultrasensitive detection of disease biomarkers with the naked eye

Nature Nanotechnology volume 7, pages 821–824 (2012)Cite this article

Subjects

Abstract

In resource-constrained countries, affordable methodologies for the detection of disease biomarkers at ultralow concentrations can potentially improve the standard of living1,2. However, current strategies for ultrasensitive detection often require sophisticated instruments that may not be available in laboratories with fewer resources3,4,5,6,7,8,9,10,11. Here, we circumvent this problem by introducing a signal generation mechanism for biosensing that enables the detection of a few molecules of analyte with the naked eye. The enzyme label of an enzyme-linked immunosorbent assay (ELISA) controls the growth of gold nanoparticles and generates coloured solutions with distinct tonality when the analyte is present. Prostate specific antigen (PSA) and HIV-1 capsid antigen p24 were detected in whole serum at the ultralow concentration of 1 × 10−18 g ml−1. p24 was also detected with the naked eye in the sera of HIV-infected patients showing viral loads undetectable by a gold standard nucleic acid-based test.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Schematic representation of the sandwich ELISA format used here and two possible signal generation mechanisms.
Figure 2: Generation of coloured solutions for detection with the naked eye.
Figure 3: Generation of nanoparticle solutions with different colours depends on the concentration of hydrogen peroxide.
Figure 4: Naked-eye detection of serum proteins with plasmonic ELISA.
Figure 5: Detection of p24 in sera from donors with different viral loads.

Similar content being viewed by others

References

  1. Patton, J. C., Coovadia, A. H., Meyers, T. M. & Sherman, G. G. Evaluation of the ultrasensitive human immunodeficiency virus type (HIV-1) p24 antigen assay performed on dried blood spots for diagnosis of HIV-1 infection in infants. Clin. Vaccine Immunol. 15, 388–391 (2008).

    Article  CAS  Google Scholar 

  2. Tang, S. & Hewlett, I. Nanoparticle-based immunoassays for sensitive and early detection of HIV-1 capsid (p24) antigen. J. Infect. Dis. 201, S59–S64 (2010).

    Article  CAS  Google Scholar 

  3. Rodriguez-Lorenzo, L., de la Rica, R., Alvarez-Puebla, R., Liz-Marzan, L. M. & Stevens, M. M. Plasmonic nanosensors with inverse sensitivity by means of enzyme-guided crystal growth. Nature Mater. 11, 604–607 (2012).

    Article  CAS  Google Scholar 

  4. Nam, J. M., Thaxton, C. S. & Mirkin, C. A. Nanoparticle-based bio-bar codes for the ultrasensitive detection of proteins. Science 301, 1884–1886 (2003).

    Article  CAS  Google Scholar 

  5. Rissin, D. M. et al. Single-molecule enzyme-linked immunosorbent assay detects serum proteins at subfemtomolar concentrations. Nature Biotechnol. 28, 595–599 (2010).

    Article  CAS  Google Scholar 

  6. Tabakman, M. N. et al. Plasmonic substrates for multiplexed protein microarrays with femtomolar sensitivity and broad dynamic range. Nature Commun. 2, 466 (2011).

    Article  Google Scholar 

  7. Laromaine, A., Koh, L. L., Murugesan, M., Ulijn, R. V. & Stevens, M. M. Protease-triggered dispersion of nanoparticle assemblies. J. Am. Chem. Soc. 129, 4156–4157 (2007).

    Article  CAS  Google Scholar 

  8. De la Rica, R., Baldi, A., Fernandez-Sanchez, C. & Matsui, H. Single-cell pathogen detection with a reverse-phase immunoassay on impedimetric transducers. Anal. Chem. 81, 7732–7736 (2009).

    Article  CAS  Google Scholar 

  9. De la Rica, R., Fratila, R. M., Szarpak, A., Huskens, J. & Velders, A. H. Multivalent nanoparticle networks as ultrasensitive enzyme sensors. Angew. Chem. Int. Ed. 50, 5703–5706 (2011).

    Google Scholar 

  10. Rodriguez-Lorenzo, L. et al. Zeptomol detection through controlled ultrasensitive surface-enhanced Raman scattering. J. Am. Chem. Soc. 131, 4616–4618 (2009).

    Article  CAS  Google Scholar 

  11. Chen, S., Svedendahl, M., van Duyne, R. P. & Käll, M. Plasmon-enhanced colorimetric ELISA with single molecule sensitivity. Nano Lett. 11, 1826–1830 (2011).

    Article  CAS  Google Scholar 

  12. Kowalczyk, B., Walker, D. A., Soh, S. & Grzybowski, B. A. Nanoparticle supracrystals and layered supracrystals as chemical amplifiers. Angew. Chem. Int. Ed. 49, 5737–5741 (2009).

    Article  Google Scholar 

  13. Aili, D., Selegard, R., Baltzer, L., Enander, K. & Liedberg, B. Colorimetric protein sensing by controlled assembly of gold nanoparticles functionalized with synthetic receptors. Small 5, 2445–2452 (2009).

    Article  CAS  Google Scholar 

  14. Qu, W., Liu, Y., Liu, D., Wang, Z. & Jiang, X. Copper-mediated amplification allows readout of immunoassays by the naked eye. Angew. Chem. Int. Ed. 50, 3442–3445 (2011).

    Article  CAS  Google Scholar 

  15. Engelbrekt, C. et al. Green synthesis of gold nanoparticles with starch–glucose and application in bioelectrochemistry. J. Mater. Chem. 19, 7839–7847 (2009).

    Article  CAS  Google Scholar 

  16. Thaxton, C. S. et al. Nanoparticle-based bio-barcode assay redefines ‘undetectable’ PSA and biochemical recurrence after radical prostatectomy. Proc. Natl Acad. Sci. USA 106, 18437–18442 (2009).

    Article  CAS  Google Scholar 

  17. De la Rica, R. & Matsui, H. Urease as a nanoreactor for growing crystalline ZnO nanoshells at room temperature. Angew. Chem. Int. Ed. 47, 5415–5417 (2008).

    Article  CAS  Google Scholar 

  18. Pejoux, C., de la Rica, R. & Matsui, H. Biomimetic crystallization of sulfide semiconductor nanoparticles in aqueous solution. Small 6, 999–1002 (2010).

    Article  CAS  Google Scholar 

  19. De la Rica, R., Fabijanic, K. I., Baldi, A. & Matsui, H. Biomimetic crystallization nanolithography: simultaneous nanopatterning and crystallization. Angew. Chem. Int. Ed. 49, 1447–1450 (2010).

    Article  CAS  Google Scholar 

  20. Fan, R. et al. Integrated barcode chips for rapid, multiplexed analysis of proteins in microliter quantities of blood. Nature Biotechnol. 26, 1373–1378 (2008).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors thank G. S. Cooke and S. Kaye (Faculty of Medicine, Imperial College London) for the kind gift of human samples containing different viral loads. M.M.S. thanks the Engineering and Physical Sciences Research Council (EPSRC) and the European Research Council (ERC) starting investigator grant ‘Naturale’ for funding. This research was supported by a Marie Curie Intra European Fellowship within the 7th European Community Framework Programme (R.R.). The monoclonal antibody to HIV-1 p24 from S. Zolla-Pazner was provided by the Centre for AIDS Reagents, National Institute for Biological Standards and Control (NIBSC, a centre of the UK Health Protection Agency).

Author information

Authors and Affiliations

  1. Department of Materials, Department of Bioengineering and Institute for Biomedical Engineering, Imperial College London, Exhibition Road, London, SW7 2AZ, UK

    Roberto de la Rica & Molly M. Stevens

Authors
  1. Roberto de la Rica
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Molly M. Stevens
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

R.R. elaborated the concept, designed and performed experiments, and wrote the paper. M.M.S. supervised the project, participated in scientific discussions, and revised the paper.

Corresponding authors

Correspondence to Roberto de la Rica or Molly M. Stevens.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary information

Supplementary information (PDF 14350 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

de la Rica, R., Stevens, M. Plasmonic ELISA for the ultrasensitive detection of disease biomarkers with the naked eye. Nature Nanotech 7, 821–824 (2012). https://doi.org/10.1038/nnano.2012.186

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nnano.2012.186

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing