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Comparing assays via the resolution of molecular concentration

Nature Biomedical Engineering volume 6, pages 227–231 (2022)Cite this article

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The assessment of the performance of molecular assays would benefit from quantifying the resolution of molecular concentration — a new metric that we propose and evaluate here.

The limit of detection (LOD) is a primary metric for assessing the performance of molecular assays. The LOD — the lowest concentration at which a signal can be statistically differentiated from the background signal1 (Box 1) — is useful for assays intended to determine the presence or absence of an analyte. However, the LOD is less useful for assays intended to discriminate changes in analyte concentration, such as assays for monitoring glucose in patients with diabetes, whose clinical care is guided by particular glucose-concentration ranges and thresholds. The performance of such ‘continuous assays’ would be better described by a quantitative metric of discriminatory resolution — for example, whether the assay can differentiate between analyte concentrations of 1 pM and 10 pM. No such standardized metric exists. In this Comment, we propose the ‘resolution of molecular concentration’ (RMC) as a simple methodology to quantify the resolution of molecular assays and to compare assay performance (Box 2). We also raise awareness of the limitations of the LOD and other typically used metrics for evaluating the performance of molecular assays.

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Fig. 1: Resolution of molecular concentration.
Fig. 2: Comparison of the performance of three commercial ELISA tests for CRP.
Fig. 3: Comparison of the performance of four point-of-care assays for the monitoring of circulating concentrations of a therapeutic monoclonal antibody.
Fig. 4: Dependence of the RMC on the parameters of a linear calibration curve.

References

  1. Long, G. L. & Winefordner, J. D. Anal. Chem. 55, 712A–724A (1983).

    Article  CAS  Google Scholar 

  2. Liang, T. et al. Adv. Funct. Mater. 29, 1903026 (2019).

    Article  CAS  Google Scholar 

  3. Armbruster, D. A. & Pry, T. Clin. Biochem. Rev. 29 (Suppl. 1), S49–S52 (2008).

    PubMed  PubMed Central  Google Scholar 

  4. Chesher, D. Clin. Biochem. Rev. 29 (Suppl. 1), S23–S26 (2008).

    PubMed  PubMed Central  Google Scholar 

  5. Currie, L. A. Anal. Chem. 40, 586–593 (1968).

    Article  CAS  Google Scholar 

  6. Currie, L. A. Anal. Chim. Acta 391, 105–126 (1999).

    Article  CAS  Google Scholar 

  7. Mandel, J. & Stiehler, R. D. J. Res. Natl Bur. Stand. 53, 155–159 (1954).

    Article  Google Scholar 

  8. Wilson, D. H. et al. Clin. Chem. 57, 1712–1721 (2011).

    Article  CAS  Google Scholar 

  9. Jarolim, P. et al. Clin. Chem. 61, 1283–1291 (2015).

    Article  CAS  Google Scholar 

  10. Pardue, H. L. Clin. Chem. 43, 1831–1837 (1997).

    Article  CAS  Google Scholar 

  11. Ridker, P. M. et al. N. Engl. J. Med. 347, 1557–1565 (2002).

    Article  CAS  Google Scholar 

  12. Eisenhardt, S. U. et al. Circ. Res. 105, 128–137 (2009).

    Article  CAS  Google Scholar 

  13. Póvoa, P. Intensive Care Med. 28, 235–243 (2002).

    Article  Google Scholar 

  14. Akin, L. et al. Horm. Res. Paediatr. 74, 72–75 (2010).

    Article  CAS  Google Scholar 

  15. Muller, P. Y. & Milton, M. N. Nat. Rev. Drug Discov. 11, 751–761 (2012).

    Article  CAS  Google Scholar 

  16. Fischer, S. K. et al. Bioanalysis 12, 1449–1458 (2020).

    Article  CAS  Google Scholar 

  17. Findlay, J. W. A. & Dillard, R. F. AAPS J. 9, E260–E267 (2007).

    Article  Google Scholar 

  18. Borysiak, M. D., Thompson, M. J. & Posner, J. D. Lab Chip 16, 1293–1313 (2016).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank M. Tirlea and M. Sklar of the Stanford Statistics Group for helpful discussions. This work was supported by the Chan–Zuckerberg Biohub investigator program, the National Institutes of Health (NIH, OT2OD025342) and the Wu Tsai Neurosciences Institute at Stanford University.

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Authors and Affiliations

  1. Department of Chemical Engineering, Stanford University, Stanford, CA, USA

    Brandon D. Wilson

  2. Department of Radiology, Stanford University, Stanford, CA, USA

    Brandon D. Wilson, Michael Eisenstein & H. Tom Soh

  3. Department of Electrical Engineering, Stanford University, Stanford, CA, USA

    Michael Eisenstein & H. Tom Soh

  4. Chan Zuckerberg Biohub, San Francisco, CA, USA

    H. Tom Soh

Authors
  1. Brandon D. Wilson
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  2. Michael Eisenstein
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  3. H. Tom Soh
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Contributions

B.D.W. and H.T.S. conceived the project. B.D.W. developed the model and analysed the data. B.D.W., M.E. and H.T.S. wrote the manuscript.

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Correspondence to H. Tom Soh.

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The authors declare no competing interests.

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Nature Biomedical Engineering thanks David Duffy and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Wilson, B.D., Eisenstein, M. & Soh, H.T. Comparing assays via the resolution of molecular concentration. Nat. Biomed. Eng 6, 227–231 (2022). https://doi.org/10.1038/s41551-021-00832-8

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