Advanced

Quantification of mitochondrial DNA copy number in suspected cancer patients by a well optimized ddPCR method

Memon, Ashfaque A. LU ; Zöller, Bengt LU ; Hedelius, Anna LU ; Wang, Xiao LU ; Stenman, Emelie LU ; Sundquist, Jan LU and Sundquist, Kristina LU (2017) In Biomolecular Detection and Quantification 13. p.32-39
Abstract

Changes in mitochondrial DNA (mtDNA) content is a useful clinical biomarker for various diseases, however results are controversial as several analytical factors can affect measurement of mtDNA. MtDNA is often quantified by taking ratio between a target mitochondrial gene and a reference nuclear gene (mtDNA/nDNA) using quantitative real time PCR often on two separate experiments. It measures relative levels by using external calibrator which may not be comparable across laboratories. We have developed and optimized a droplet digital PCR (ddPCR) based method for quantification of absolute copy number of both mtDNA and nDNA gene in whole blood. Finally, the role of mtDNA in suspected cancer patients referred to a cancer diagnostic center... (More)

Changes in mitochondrial DNA (mtDNA) content is a useful clinical biomarker for various diseases, however results are controversial as several analytical factors can affect measurement of mtDNA. MtDNA is often quantified by taking ratio between a target mitochondrial gene and a reference nuclear gene (mtDNA/nDNA) using quantitative real time PCR often on two separate experiments. It measures relative levels by using external calibrator which may not be comparable across laboratories. We have developed and optimized a droplet digital PCR (ddPCR) based method for quantification of absolute copy number of both mtDNA and nDNA gene in whole blood. Finally, the role of mtDNA in suspected cancer patients referred to a cancer diagnostic center was investigated.Analytical factors which can result in false quantification of mtDNA have been optimized and both target and reference have been quantified simultaneously with intra- and inter-assay coefficient variances as 3.1% and 4.2% respectively. Quantification of mtDNA show that compared to controls, solid tumors (but not hematologic malignancies) and other diseases had significantly lower copy number of mtDNA. Higher mtDNA (highest quartile) was associated with a significantly lower risk of both solid tumors and other diseases, independent of age and sex. Receiver operating curve demonstrated that mtDNA levels could differentiate controls from patients with solid tumors and other diseases.Quantification of mtDNA by a well optimized ddPCR method showed that its depletion may be a hallmark of general illness and can be used to stratify healthy individuals from patients diagnosed with cancer and other chronic diseases.

(Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Biomarker, Cancer, DdPCR, Droplet digital PCR, Mitochondrial DNA, MtDNA, Nuclear DNA
in
Biomolecular Detection and Quantification
volume
13
pages
32 - 39
publisher
Elsevier GmbH
external identifiers
  • scopus:85028603303
DOI
10.1016/j.bdq.2017.08.001
language
English
LU publication?
yes
id
9b35fa9e-5514-4c1d-8c6c-83ee3a7bb6cc
date added to LUP
2017-09-27 13:50:54
date last changed
2018-04-15 04:47:26
@article{9b35fa9e-5514-4c1d-8c6c-83ee3a7bb6cc,
  abstract     = {<p>Changes in mitochondrial DNA (mtDNA) content is a useful clinical biomarker for various diseases, however results are controversial as several analytical factors can affect measurement of mtDNA. MtDNA is often quantified by taking ratio between a target mitochondrial gene and a reference nuclear gene (mtDNA/nDNA) using quantitative real time PCR often on two separate experiments. It measures relative levels by using external calibrator which may not be comparable across laboratories. We have developed and optimized a droplet digital PCR (ddPCR) based method for quantification of absolute copy number of both mtDNA and nDNA gene in whole blood. Finally, the role of mtDNA in suspected cancer patients referred to a cancer diagnostic center was investigated.Analytical factors which can result in false quantification of mtDNA have been optimized and both target and reference have been quantified simultaneously with intra- and inter-assay coefficient variances as 3.1% and 4.2% respectively. Quantification of mtDNA show that compared to controls, solid tumors (but not hematologic malignancies) and other diseases had significantly lower copy number of mtDNA. Higher mtDNA (highest quartile) was associated with a significantly lower risk of both solid tumors and other diseases, independent of age and sex. Receiver operating curve demonstrated that mtDNA levels could differentiate controls from patients with solid tumors and other diseases.Quantification of mtDNA by a well optimized ddPCR method showed that its depletion may be a hallmark of general illness and can be used to stratify healthy individuals from patients diagnosed with cancer and other chronic diseases.</p>},
  author       = {Memon, Ashfaque A. and Zöller, Bengt and Hedelius, Anna and Wang, Xiao and Stenman, Emelie and Sundquist, Jan and Sundquist, Kristina},
  keyword      = {Biomarker,Cancer,DdPCR,Droplet digital PCR,Mitochondrial DNA,MtDNA,Nuclear DNA},
  language     = {eng},
  month        = {08},
  pages        = {32--39},
  publisher    = {Elsevier GmbH},
  series       = {Biomolecular Detection and Quantification},
  title        = {Quantification of mitochondrial DNA copy number in suspected cancer patients by a well optimized ddPCR method},
  url          = {http://dx.doi.org/10.1016/j.bdq.2017.08.001},
  volume       = {13},
  year         = {2017},
}