• Quantification of Mitochondrial DNA in Human Cells Using Real-Time PCR With Fluorogenic Reporters (TuPeB4509)
  Authored by D.M. Paulsen, B.J. Hill, G. McComsey, L.L. Ross
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Denise Paulson from GlaxoSmithKline and colleagues reported on their effort to develop a rapid and less complicated tool for assessing the status of mitochondrial DNA (mtDNA) in patients taking NRTIs for HIV infection.

Use of the real-time PCR assay with fluorogenic reporters in fat cells found a 50 percent reduction in mtDNA in fat cells from patients experiencing lipodystrophy compared to patients without lipodystrophy. When peripheral blood mononuclear cells (PBMCs) were assessed, only modest differences were noted between PBMCs from patients with lipodystrophy (n = 16; 136 genomes/cell), HIV negative controls (n = 51; 156 genomes/cell), HIV-positive patients on NRTI therapy more than six months without lipodystrophy (n = 10; 173 genomes/cell), or HIV-positive patients who were treatment naive (n = 10; 188 genomes/cell). It is unclear whether 10 percent less mtDNA in the PBMCs from patients with lipoatrophy compared to patients without lipoatrophy is significant (no statistical analysis was performed). Another poster (TuPeB4500) using a PCR-based technique also found no difference between mtDNA levels from patients on NRTI-based therapy and matched normal controls. One observer at the poster remarked that platelets can contaminate the PBMC fraction and can contribute enough mitochondria to throw the results off. More work is needed to find a blood-based assay to detect early mitochondrial toxicity. Other observers commented that PMNs may be a worthwhile fraction to study as well.