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

S Damodara, J Arora, PC Liaw, AE Fox-Robichaud… - Microchimica Acta, 2022 - Springer
Microchimica Acta, 2022Springer
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 …
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.
Graphical abstract
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