Digital quantification of random mitochondrial DNA deletions

Description:

Digital Deletion Detection and QuantiSize

This method allows for measurement of rare deletions, while concurrently gathering information about size and homogeneity of the unknown template.

Dr. Bielas has developed a sensitive, quantitative method to detect rare mutations in genomic and mitochondrial DNA. The main strategy is a three-step process starting with targeted enrichment for deletions, followed by amplification, and then analysis for quantification or characterization. This method shows improvements in specificity, sensitivity, and accuracy over other available methods. In addition, the lab observed a linear correlation between the fluorescence amplitude produced in droplet digital PCR (ddPCR) and the length of the amplified DNA molecule, which they named QuantiSize. QuantiSize is based off of the ddPCR absolute quantification system and adds in the ability to determine the size of a target DNA. This allows for accurate preparation of NGS libraries, while avoiding limitations of other quantification systems.

<ul>

<li>Detection of rare mutations in plurality of nucleic acid molecule</li>

<li>Determination of nucleic acid size using ddPCR</li>

<li>Development of assays using mtDNA deletions as biomarkers for diagnostic or early detection</li>

</ul>

<ul>

<li>Measures rare deletions at frequencies as low as 1 deletion per 100 million genomes</li>

<li>Accurate and precise method with high sensitivity</li>

<li>Saves time by measuring length and size concurrently</li>

</ul>

<ul>

<li>Jason Bielas, PhD, Public Health and Human Biology Divisions</li>