Mitochondrial DNA Deletion Analysis: A Comparison of PCR Quantitative Methods

The role of mitochondrial DNA (mtDNA) deletions in aging and in neurodegenerative diseases is often determined by measuring the amount of deleted mtDNA in the affected tissue. Upon examining brain autopsy tissue from a 59 year old individual with lung cancer we determined by serial dilution PCR and...

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Bibliographic Details
Published in:Biochemical and biophysical research communications 1995-02, Vol.207 (2), p.839-847
Main Authors: Hamblet, N.S., Castora, F.J.
Format: Article
Language:English
Subjects:
Base Sequence
Brain - metabolism
Brain - pathology
Caudate Nucleus - metabolism
Caudate Nucleus - pathology
DNA Primers
DNA, Mitochondrial - analysis
DNA, Mitochondrial - genetics
Humans
Kinetics
Lung Neoplasms - genetics
Lung Neoplasms - pathology
Middle Aged
Molecular Sequence Data
Parietal Lobe - metabolism
Parietal Lobe - pathology
Polymerase Chain Reaction - methods
Reference Values
Sequence Deletion
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Summary:The role of mitochondrial DNA (mtDNA) deletions in aging and in neurodegenerative diseases is often determined by measuring the amount of deleted mtDNA in the affected tissue. Upon examining brain autopsy tissue from a 59 year old individual with lung cancer we determined by serial dilution PCR and kinetic PCR that a greater ratio of deleted mtDNA was present in the caudate than in the parietal cortex. However, the magnitude difference for these two brain regions appeared to be technique dependent; by serial dilution PCR the caudate had 10 times more deleted mtDNA than the parietal cortex (0.0141 vs 0.0014) whereas kinetic PCR yielded a 4-fold difference (0.1258 vs 0.0316). These results indicate that although it is valid to compare the amount of deleted mtDNA in normal and diseased tissue and draw conclusions based on relative comparisons within one study, greater caution should be exercised when comparing absolute values from studies using different measurement techniques.
ISSN:0006-291X
1090-2104
DOI:10.1006/bbrc.1995.1262