Mb. Coulter-mackie et al., A protocol for detection of mitochondrial DNA deletions: Characterization of a novel deletion, CLIN BIOCH, 31(8), 1998, pp. 627-632
Objectives: To develop a protocol capable of identifying deletions in mitoc
hondrial DNA and use it to identify the breakpoints of a mtDNA deletion in
a patient with chronic progressive external ophthalmoplegia (CPEO).
Design and Methods: Deletions in mtDNA were identified by a combination of
long range PGR and Southern blotting. The precise breakpoints were determin
ed by automated DNA sequencing.
Results: A series of DNA samples from patients with suspected mitochondrial
disease was subjected to a protocol, which combines long range PGR and Sou
thern blotting. We found a unique deletion in a patient with CPEO and we id
entified the precise location of this deletion through DNA sequencing.
Conclusions: Long range PCR has the advantages of speed, minimal sample req
uirements, and sensitivity. Southern blotting is better able to evaluate he
teroplasmy and detect duplications. We suggest a protocol that enables us t
o identify precisely the breakpoints in a unique mutation of mtDNA in a pat
ient with CPEO. (C) 1998 The Canadian Society of Clinical Chemists.