Emulsion digital PCR
Ended Project Grant Awarded Last action
- Award ID
- R44AI174618
- Recipient
-
Dplexbio Inc. UEI LKU9MBA8JHA7
- Awarding agency
-
National Institute of Allergy and Infectious Diseases
National Institutes of Health, Department of Health and Human Services
- Obligated
- $2,170,226
Award facts
- Assistance listing
- 93.242 Mental Health Research Grants
- Completion date
- Funding agency
- National Institute of Mental Health National Institutes of Health, Department of Health and Human Services
- Place of performance
- Washington, USA
Award details come from USAspending.gov.
Description
- Abstract
- This proposed project aims to develop a new type of digital PCR platform which eliminates the need for uniform volumes and thereby reduces the complexity, cost, and run time of digital PCR while increasing its dynamic range. The result is a higher performance digital PCR system that matches the simplicity, speed, and low per-assay cost of real-time PCR. Real-time PCR maintains great popularity in clinical diagnostics, but digital PCR is superior to real-time PCR because IT provides absolute quantitation, greater accuracy at low concentrations, and greater reproducibility. However, because IT was hypothesized that uniform volumes are required for digital PCR quantitation, current digital PCR platforms require precise microfluidic chips and control, which result in low throughput and high per-assay costs. Here we propose to address these limitations by developing a digital PCR system that employs variable volume droplets created simply by shaking a sample along with PCR reagents and an oil/emulsifier mix to create an emulsion. PCR is performed, and droplets in the emulsion are imaged and sized. Droplets with one or more copies of a nucleic acid are identified, and nucleic acid concentration is determined. We call this edpcr (emulsion digital PCR). We characterized this edcpr method via computational simulations and validated IT experimentally. Simulations were used to investigate the dependence of droplet occupancy on analyte concentration and droplet size distribution, and to estimate the accuracy of the measured concentration in the presence of errors in measurement of droplet volume. Simulations also provided an estimate of dynamic range for a given droplet size distribution and statistical power. The method was validated experimentally in terms of accuracy, precision, and dynamic range, and to rule out potential sources of biased error, such as droplet shrinking and droplet fusion during PCR. In the proposed work, we will develop a commercial edpcr platform that will offer the superior performance of digital PCR while providing the high-throughput (e.g. 384 well plate operation) and low per-assay cost of real-time PCR, which we believe will remove the barriers to the widespread use of digital PCR in clinical assays, point-of-care settings, and large-scale testing.
Transactions
| Action | Obligated | Date |
|---|---|---|
| Amendment | $0 | |
| Amendment | $192,866 | |
| Amendment | $494,340 | |
| Amendment | $494,340 | |
| Amendment | $494,340 | |
| Original award | $494,340 |
Quick answers
Who received award R44AI174618?
R44AI174618 was awarded to Dplexbio Inc. by National Institute of Allergy and Infectious Diseases (National Institutes of Health) on March 17, 2023.
How much is award R44AI174618 worth?
$2,170,226 has been obligated on R44AI174618.
Which agency awarded R44AI174618?
National Institute of Allergy and Infectious Diseases (National Institutes of Health) awarded R44AI174618, with funding from National Institute of Mental Health (National Institutes of Health).
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