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Single-step measurement of cell-free DNA for sepsis prognosis using a thread-based microfluidic device

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Abstract

Cell-free DNA (cfDNA) content in plasma has been studied as a biomarker for sepsis. Recent publications show that the cfDNA content in sepsis patients entering intensive care unit who were likely to survive had a total cfDNA concentration of 1.16 ± 0.13 μg/mL compared to 4.65 ± 0.48 μg/mL of non-survivors. Current methods for measuring cfDNA content in plasma were designed to amplify and measure low concentrations of specific DNA, making them unsuitable for low-cost measurement of total cfDNA content in plasma. Here, we have developed a point of care (POC) device that uses a thread silicone device as a medium to store a fluorescent dye which eliminates the need for preparatory steps, external aliquoting and dispensing of reagents, preconcentration, and external mixing while reducing the detection cost. The device was paired with a portable imaging system with an excitation filter at 472 ± 10 nm and an emission filter of 520 ± 10 nm that can be operated with just 100 mA current supply. The device was demonstrated for use in the quantification of buffered cfDNA samples in a range 1–6 μg/mL with a sensitivity of 5.72 AU/μg/mL and with cfDNA spiked in plasma with a range of 1–3 μg/mL and a sensitivity of 5.43 AU/μg/mL. The results showed that the device could be used as a low-cost, rapid, and portable POC device for differentiating between survivors and non-survivors of sepsis within 20 min.

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Acknowledgements

P.R.S. also acknowledges support from the Canada Research Chairs Program as well as the Discovery Accelerator Supplement grant.

Funding

This work was supported by the Natural Sciences and Engineering Research Council of Canada (NSERC) and Canadian Institutes for Health Research (CIHR) through the Collaborative Health Research Program.

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Authors and Affiliations

  1. Department of Mechanical Engineering, McMaster University, Hamilton, Canada

    Sreekant Damodara & P. Ravi Selvaganapathy

  2. Department of Medicine, McMaster University, Hamilton, Canada

    Jaskirat Arora, Patricia C. Liaw & Alison E. Fox-Robichaud

Authors
  1. Sreekant Damodara
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  2. Jaskirat Arora
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  3. Patricia C. Liaw
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  4. Alison E. Fox-Robichaud
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  5. P. Ravi Selvaganapathy
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Consortia

the Canadian Critical Care Translational Biology Group

Contributions

S.D. and P.R.S. formulated the concept and experimental protocols with inputs from A.E.F. J.A. and P.C.L. provided cfDNA samples and provided inputs on biomarkers. S.D. wrote the paper with revisions and inputs from P.R.S., A.E.F., P.C.L., and C.C.T.B.G.

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Correspondence to P. Ravi Selvaganapathy.

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The authors declare no competing interests.

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Damodara, S., Arora, J., Liaw, P.C. et al. Single-step measurement of cell-free DNA for sepsis prognosis using a thread-based microfluidic device. Microchim Acta 189, 146 (2022). https://doi.org/10.1007/s00604-022-05245-1

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  • DOI: https://doi.org/10.1007/s00604-022-05245-1

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