Corporate Banner
Satellite Banner
qPCR
Scientific Community
 
Become a Member | Sign in
Home>News>This Article
  News
Return

Accurate Detection of Extremely Rare Mitochondrial DNA Deletions Associated with Aging

Published: Thursday, September 05, 2013
Last Updated: Thursday, September 05, 2013
Bookmark and Share
The study published in Aging Cell identifies a new tool to accurately analyze extremely rare mitochondrial DNA deletions associated with a range of diseases and disorders as well as aging.

This approach, which relies on Droplet Digital PCR (ddPCR™) technology, will help researchers explore mitochondrial DNA (mtDNA) deletions as potential disease biomarkers.

The accumulation of mtDNA mutations is associated with aging, neuromuscular disorders, and cancer. However, methods to probe the underlying mechanisms behind this mutagenesis have been limited by their inability to accurately quantify and characterize new deletion events, which may occur at a frequency as low as one deletion event per 100 million mitochondrial genomes in normal tissue. To address these limitations, researchers at the Seattle, Washington-based Fred Hutchinson Cancer Research Center developed a ddPCR-based assay known as "Digital Deletion Detection" (3D) that allows for the high-resolution analysis of these rare deletions.

"It is incredibly difficult to study mtDNA mutations, let alone deletions, within the genome," said Dr. Jason Bielas, Assistant Member of the Public Health Sciences Division at the Fred Hutchinson Cancer Research Center and lead author of the study. "Our 3D assay shows significant improvement in specificity, sensitivity, and accuracy over conventional methods such as those that rely on real-time PCR."

Bielas added, "The increase in throughput afforded by droplet digital PCR shortened the analysis of deletion events to days compared to months using previous digital PCR methods. Without the technology, we could not have made this discovery."

At the center of the study was Bio-Rad Laboratories' QX100™ ddPCR system. Using the QX100 system, Bielas and his team analyzed eight billion human brain mtDNA genomes and identified more than 100,000 genomes with a deletion. They discovered that, contrary to popular belief, the majority of the increase in mtDNA deletions was not caused by new deletions but rather by the expansion of previous deletions. They hypothesized that the expansion of pre-existing mutations should be considered as the primary factor contributing to age-related accumulation of mtDNA deletions.

How the 3D Assay Works
3D is a novel three-step process that includes enrichment for deletion-bearing molecules, single-molecule partitioning of genomes into droplets for direct quantification via ddPCR, and breakpoint characterization using next-generation sequencing.

Once the enrichment process is completed using methods previously developed by Bielas and colleagues, the concentration of molecules within the droplets is adjusted by using the QX100 system so that the majority of droplets contain no mutant genomes while a small fraction contain only one. This process allows each deletion to be amplified without bias and without introducing the artifacts that are common in qPCR.

Following amplification, deletions can be analyzed using ddPCR to determine the absolute concentration of mutated molecules. Using the relationship between droplet fluorescence and amplicon size, Bielas and his team were able to characterize the size and complexity (whether they were a result of a few clonal expansions or a large collection of random deletions) of rare mitochondrial deletions in human brain samples.

The 3D assay provides an important new tool that will allow researchers to better study the mechanisms of deletion formation and expansion, and their role in aging. Droplet digital PCR's high throughput and increased sensitivity will also allow Bielas' lab to target other low-level disease-causing mtDNA deletions in skeletal muscle, brain tissue, and blood.


Further Information

Join For Free

Access to this exclusive content is for Technology Networks Premium members only.

Join Technology Networks Premium for free access to:

  • Exclusive articles
  • Presentations from international conferences
  • Over 3,300+ scientific posters on ePosters
  • More than 4,800+ scientific videos on LabTube
  • 35 community eNewsletters


Sign In



Forgotten your details? Click Here
If you are not a member you can join here

*Please note: By logging into TechnologyNetworks.com you agree to accept the use of cookies. To find out more about the cookies we use and how to delete them, see our privacy policy.

Related Content

Assessing the Effectiveness of Genome-Editing Technologies
Researchers have developed a cost-effective and rapid method for assessing edits generated by CRISPR-Cas9 and other genome-editing technologies.
Thursday, June 30, 2016
Bio-Rad QX200 ddPCR System Recieves CE IVD Mark
The first digital PCR instrument with the CE IVD mark.
Monday, February 29, 2016
Presentations at the AACR Annual Meeting to Highlight Advances in Cancer Research Made Possible by Bio-Rad’s Droplet Digital™ PCR Technology
Tailoring Treatments and Tracking Mutations with Liquid Biopsies possible with ddPCR™ technology
Wednesday, April 22, 2015
Bio-Rad Reports Fourth-Quarter and Full-Year 2014 Financial Results
Company has introduced its S3e™ Cell Sorter during the quarter.
Saturday, March 07, 2015
Accurate Determination of Copy Number States for Multiallelic Copy Number Variations
Researchers use next-generation sequencing (NGS) and Bio-Rad’s Droplet Digital PCR (ddPCR™) technology to solve the technical challenge.
Tuesday, March 03, 2015
Awards for Bio-Rad's Protein Expression Products and Genomic Workflow Solutions
BioInformatics LLC has recognized Bio-Rad with two prestigious Life Science Industry Awards for “Best Protein Expression & Analysis Products” and “Best Workflow Solutions — Genomics.”
Monday, January 19, 2015
Bio-Rad Acquires Sequencing Technology Company GnuBIO
GnuBIO is a privately-held life sciences company that has developed a droplet-based DNA sequencing technology.
Friday, April 11, 2014
Droplet Digital PCR Enables Measurement of Potential Cancer Survival Biomarker
Study paves the way for further research into the role of TIL quantification in immunotherapy and as a cancer survival predictor.
Tuesday, December 10, 2013
A Brief RT-qPCR “Field Guide” for MIQE Adherence
Bio-Rad’s Sean Taylor and Eli Mrkusich have published a practical guide for MIQE compliance.
Monday, December 09, 2013
Researchers Develop Rapid, Cost-Effective Early Detection Method for Organ Transplant Injury
Chronix Biomedical and transplant expert Prof. Oellerich use Droplet Digital™ PCR to quantify early rejection biomarker.
Thursday, August 29, 2013
Accurate Quantification of NGS Libraries
A study has found that Droplet Digital PCR (ddPCR™) can be used as an accurate and precise method for quality control of NGS libraries.
Tuesday, August 20, 2013
New Biomarker Could Reveal Alzheimer's Disease Years Before Onset
A new study has reported the identification of what may be the earliest known biomarker associated with the risk of developing Alzheimer's disease (AD).
Wednesday, August 14, 2013
Droplet Digital™ PCR Works for GMO Quantification
A study published in the journal PLOS ONE has found that Droplet Digital PCR technology is suitable for routine analysis of genetically modified organisms in food, feed and seeds.
Wednesday, May 08, 2013
Bio-Rad Receives Award for QX100 Droplet Digital PCR System
Company receives Frost & Sullivan 2012 North American Laboratory Researchers’ Choice: Future market leader of digital PCR technology award.
Thursday, March 14, 2013
Bio-Rad Receives North American Market Penetration Award
The award honors the company that has grown market share at the fastest rate in its industry, as measured by revenues or units sold.
Tuesday, November 20, 2012
Scientific News
Lab-Tested Diagnosis Needed When Treating Persistent Diarrhea
New PCR multiplex method makes lab testing more effective.
Biomarker for Multiple Sclerosis Detection Discovered
Winthrop-University Hospital researchers discover biomarker for multiple sclerosis detection.
Scientists Link Bipolar Disorder to Unexpected Brain Region
Researchers from The Scripps Research Institute have found that gene within the brain’s striatum could be linked to biopolar disorder.
Quick, Early Test For Ebola Could Prevent Epidemics
Researchers from Princeton University are collaborating with U.S. government labs to develop a more rapid, accurate and inexpensive test for the Ebola virus with the aim of identifying infections before carriers become symptomatic and contagious.
From Super to Ultra-Resolution Microscopy
A new method pushes the frontier in imaging resolution, with the potential to distinguish individual features in single molecules.
Cancer Research UK joins forces with U.S. 'Cancer Moonshot'
Cancer Research UK and the US government’s National Cancer Institute have announced that two teams will work together to radically accelerate progress against cancer, in one of the first international collaborations inspired by US Vice President Joe Biden’s Cancer Moonshot.
Assessing the Effectiveness of Genome-Editing Technologies
Researchers have developed a cost-effective and rapid method for assessing edits generated by CRISPR-Cas9 and other genome-editing technologies.
Measuring the Abundance of Extremely Rare Mutations
Researchers from Rutgers University demonstrate the use of multiplex real-time PCR assays to measure the abundance of extremely rare mutations associated with cancer.
Harnessing Helpful Microbes
Seeking to further harness microbes’ many uses, the federal government has launched the National Microbiome Initiative (NMI) to “foster the integrated study of microbiomes across different ecosystems.”
What Makes a Good Scientist?
It’s the journey, not just the destination that counts as a scientist when conducting research.

SELECTBIO Market Reports
Go to LabTube
Go to eposters
 
Access to the latest scientific news
Exclusive articles
Upload and share your posters on ePosters
Latest presentations and webinars
View a library of 1,800+ scientific and medical posters
3,300+ scientific and medical posters
A library of 2,500+ scientific videos on LabTube
4,800+ scientific videos
Close
Premium CrownJOIN TECHNOLOGY NETWORKS PREMIUM FOR FREE!