PCR amplification using a single cell allows the detection of the mtDNA lesion associated with Leber's hereditary optic neuropathy

Biochim Biophys Acta. 1993 Mar 24;1181(1):77-82. doi: 10.1016/0925-4439(93)90093-g.


The development of the polymerase chain reaction (PCR), which routinely can amplify specific target sequences more than one billion-fold, has made it possible to produce readily detectable amounts of DNA from a few copies of very rare sequences. We have begun a study of mitochondrial myopathies with the purpose of developing a diagnostic test using PCR to amplify appropriate mitochondrial DNA (mtDNA) target sequences from small amounts of sample. We have developed a 15-min procedure for recovering mtDNA which can be amplified by PCR to detectable levels, from as little as 30 microliters of blood or 5 microliters of amniotic fluid. We have microscopically selected HL60 cells, and have found that 28 cycles of PCR allows the detection of mitochondrial targets from a single cell. Using micromanipulation techniques, we utilized this approach to analyze mtDNA from a single cell isolated from an 8-cell stage mouse blastocyst. Finally, a single cell cultured from a patient with Leber's hereditary optic neuropathy, a mitochondrial myopathy, provided sufficient mtDNA for detection of the single base substitution that leads to loss of a restriction endonuclease recognition site for SfaNI and generation of a site for MaeIII.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cells, Cultured
  • DNA Mutational Analysis
  • DNA, Mitochondrial / blood
  • DNA, Mitochondrial / genetics*
  • Electrophoresis, Polyacrylamide Gel
  • Humans
  • Mice
  • Optic Atrophies, Hereditary / genetics*
  • Polymerase Chain Reaction / methods*


  • DNA, Mitochondrial