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Updated: Dec 15, 2025
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Methylation Specific Multiplex Droplet PCR using Polymer Droplet Generator Device for Hematological Diagnostics
Published on: June 29, 2020
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Methylation Specific Multiplex Droplet PCR using Polymer Droplet Generator Device for Hematological Diagnostics
Lidija Malic1, Abdelrahman Elmanzalawy2, Jamal Daoud3
1Life Sciences Division, National Research Council of Canada; Lidija.Malic@cnrc-nrc.gc.ca.
Journal of Visualized Experiments : Jove
|July 14, 2020
Summary
This study introduces a new method using multiplexed droplet PCR (mdPCR) to count white blood cell (WBC) types by analyzing DNA methylation. This epigenetic approach offers precise quantification for improved clinical diagnosis and prognosis.
Area of Science:
- Biochemistry
- Molecular Biology
- Genomics
Background:
- Epigenetic markers, specifically DNA methylation patterns at CpG loci, are crucial for white blood cell (WBC) subtyping.
- Accurate WBC differential counts have significant prognostic value in various diseases.
Purpose of the Study:
- To develop and detail a multiplexed droplet PCR (mdPCR) workflow for epigenetic-based WBC differential counting.
- To present a thermoplastic elastomer (TPE) microfluidic device for droplet generation in this workflow.
Main Methods:
- Utilized bisulfite-treated DNA from peripheral blood mononuclear cells (PBMCs).
- Encapsulated DNA with mdPCR reagents, primers, and fluorescent probes targeting specific CpG loci correlated with WBC sub-populations within droplets.
- Employed a multiplexed approach to interrogate multiple CpG loci simultaneously.
Main Results:
- Demonstrated a precise quantification method for WBC sub-populations based on DNA methylation analysis.
- The multiplexed droplet PCR workflow enables simultaneous analysis of numerous CpG loci, enhancing accuracy.
Conclusions:
- The described mdPCR workflow and TPE microfluidic device provide an accurate method for epigenetic-based WBC differential counting.
- This precise quantification technique has broad applications in clinical diagnosis and prognosis.
