title: Quantification of mitochondrial toxicity in HIV-infected individuals by quantitative PCR compared to flow cytometry.
abstract: BACKGROUND
  Non-invasive diagnostic assays to evaluate mitochondrial toxicity could have significant clinical utility for HIV-infected individuals on antiretroviral therapy (ART).
  
  
  METHODS
  This study compared the ratio of mitochondrial to nuclear DNA determined by quantitative polymerase chain reaction (qPCR) to the ratio of mitochondrial to nuclear-encoded proteins by flow cytometry, in peripheral blood mononuclear cells from 73 HIV-infected individuals with and without risk factors for mitochondrial toxicity.
  
  
  RESULTS
  PCR detected similar mitochondrial/nuclear DNA in HIV-infected individuals without a history of ART, and those receiving ART with lipodystrophy, lipoatrophy, or a history of suspected lactic acidosis. However, the ratio was significantly greater in ART-untreated compared to those receiving either stavudine or didanosine. In contrast, flow cytometry did not detect any differences in mitochondrial/nuclear protein (Lin et al., Cytometry B 2009;76B:181-190). There was no correlation between the assays (rho = -0.05, P = 0.65).
  
  
  CONCLUSIONS
  Assessment of the mitochondrial/nuclear DNA ratio by qPCR performed better than the mitochondrial/nuclear-encoded protein ratio by flow cytometry to detect adverse effects of nucleoside analogs on mitochondria.
authors.affiliation: Seattle Children's Research Institute, Seattle, Washington 98101, USA. agner@seattlechildrens.org
authors.forename: Thor A
authors.initials: TA
authors.lastname: Wagner
authors.forename: Chen-Han
authors.initials: CH
authors.lastname: Lin
authors.forename: Nicole H
authors.initials: NH
authors.lastname: Tobin
authors.forename: Hélène C F
authors.initials: HC
authors.lastname: Côté
authors.forename: Derek D
authors.initials: DD
authors.lastname: Sloan
authors.forename: Keith R
authors.initials: KR
authors.lastname: Jerome
authors.forename: Lisa M
authors.initials: LM
authors.lastname: Frenkel
mesh_terms: D000140:Acidosis, Lactic; D000328:Adult; D019380:Anti-HIV Agents / Q000627:therapeutic use; D023241:Antiretroviral Therapy, Highly Active; D004272:DNA, Mitochondrial / Q000032:analysis*; D016049:Didanosine / Q000627:therapeutic use; D005260:Female; D005434:Flow Cytometry; D015658:HIV Infections / Q000097:blood / Q000175:diagnosis*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000166:cytology; D008060:Lipodystrophy; D008297:Male; D008875:Middle Aged; D008928:Mitochondria / Q000473:pathology*; D024101:Mitochondrial Proteins / Q000032:analysis*; D060888:Real-Time Polymerase Chain Reaction; D018119:Stavudine / Q000627:therapeutic use
chemical_list: D019380:Anti-HIV Agents; D004272:DNA, Mitochondrial; D024101:Mitochondrial Proteins; D018119:Stavudine; D016049:Didanosine