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

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
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 2,800+ scientific posters on ePosters
  • More than 4,000+ 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 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

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
Case Study Demonstrates Importance of MIQE for qPCR Data Analysis
Published by Bio-Rad Laboratories, Inc. this case study demonstrates how real-time PCR can lead to erroneous conclusions if the key steps set out in the MIQE guidelines are not followed.
Friday, February 17, 2012
Bio-Rad Acquires QuantaLife and Digital PCR Technology
Bio-Rad purchases QuantaLife for $162 million in cash plus potential future milestone payments.
Friday, October 07, 2011
Scientific News
Ancient Viral Molecules Essential for Human Development
Genetic material from ancient viral infections is critical to human development, according to researchers at the Stanford University School of Medicine.
Measuring microRNAs in Blood to Speed Cancer Detection
A simple, ultrasensitive microRNA sensor holds promise for the design of new diagnostic strategies and, potentially, for the prognosis and treatment of pancreatic and other cancers.
Best Test to Diagnose Strangles in Horses Identified
New research by Dr. Ashley Boyle of New Bolton Center’s Equine Field Service team shows that the best method for diagnosing Strangles in horses is to take samples from a horse’s guttural pouch and analyze them using a loop-mediated amplification (LAMP) polymerase chain reaction (PCR) test.
Tardigrade's Are DNA Master Thieves
Tardigrades, nearly microscopic animals that can survive the harshest of environments, including outer space, hold the record for the animal that has the most foreign DNA.
Rapid, Portable Ebola Diagnostic
Scientists confirmed the efficiency of the novel Ebola detection method in field trials.
Detecting When Hormone Treatment for Breast Cancer Stops Working
Scientists have developed a highly sensitive blood test that can spot when breast cancers become resistant to standard hormone treatment, and have demonstrated that this test could guide further treatment.
Packaging and Unpacking of the Genome
New research improves understanding of the importance of histone replacement.
New Way to Find DNA Damage
University of Utah chemists devised a new way to detect chemical damage to DNA that sometimes leads to genetic mutations responsible for many diseases, including various cancers and neurological disorders.
How Different Treatments for Crohn's Effect the Microbiome
Different treatments for Crohn's disease in children affects their gut microbes in distinct ways, which has implications for future development of microbial-targeted therapies for these patients, according to a study led by researchers from the Perelman School of Medicine at the University of Pennsylvania.
Charting the 'Genomic Biography' of Leukemia
A new study by scientists at Dana-Farber Cancer Institute and the Broad Institute of MIT and Harvard offers a glimpse of the wealth of information that can be gleaned by combing the genome of a large collection of leukemia tissue samples.
Skyscraper Banner

Skyscraper Banner
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
2,800+ scientific and medical posters
A library of 2,500+ scientific videos on LabTube
4,000+ scientific videos