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Expanded Multiplexing on Sensor-Constrained Microfluidic Partitioning Systems.

Pavan K Kota1, Hoang-Anh Vu1, Daniel LeJeune2

  • 1Department of Bioengineering, Rice University, Houston, Texas 77005, United States.

Analytical Chemistry
|November 16, 2023
PubMed
Summary
This summary is machine-generated.

We present Sparse Poisson Recovery (SPoRe), a scalable method to multiplex analytes using microfluidic partitioning. This approach enhances digital PCR for infection diagnostics by overcoming channel limitations and enabling accurate bacterial quantification.

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Area of Science:

  • Biotechnology
  • Molecular Diagnostics
  • Microfluidics

Background:

  • Microfluidic partitioning, such as droplets or nanowells, captures analytes following a Poisson distribution.
  • Maximum Likelihood Estimation (MLE) is commonly used for inferring analyte concentration in diagnostics.
  • Existing methods face limitations in multiplexing capacity, especially in digital PCR.

Purpose of the Study:

  • To present a scalable approach for multiplexing analytes using microfluidic partitioning.
  • To generalize Maximum Likelihood Estimation (MLE) and extend the Sparse Poisson Recovery (SPoRe) algorithm.
  • To demonstrate the first in vitro application of SPoRe with droplet digital PCR (ddPCR) for infection diagnostics.

Main Methods:

  • Generalizing MLE with microfluidic partitioning and extending the Sparse Poisson Recovery (SPoRe) algorithm.
  • Implementing SPoRe with a two-channel droplet digital PCR (ddPCR) system for bacterial identification.
  • Utilizing 16S ddPCR with five nonspecific probes to barcode nine pathogen genera.

Main Results:

  • Successfully recovered bacterial concentrations from pooled droplet data, despite individual droplet ambiguity.
  • Achieved stable quantification down to approximately 200 total copies of the 16S gene per sample.
  • Demonstrated expanded multiplexing capacity for digital PCR by circumventing channel limitations.

Conclusions:

  • SPoRe offers a scalable solution for analyte multiplexing in microfluidic partitioning systems.
  • The developed framework and scaling rules can impact various biosensing applications.
  • This approach enables robust microbial DNA quantification for clinical diagnostics.