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Quantification of mitochondrial toxicity in HIV‐infected individuals by quantitative PCR compared to flow cytometry
Author(s) -
Wagner Thor A.,
Lin ChenHan,
Tobin Nicole H.,
Côté Hélène C. F.,
Sloan Derek D.,
Jerome Keith R.,
Frenkel Lisa M.
Publication year - 2012
Publication title -
cytometry part b: clinical cytometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.646
H-Index - 61
eISSN - 1552-4957
pISSN - 1552-4949
DOI - 10.1002/cyto.b.21045
Subject(s) - mitochondrial toxicity , flow cytometry , didanosine , stavudine , nuclear dna , mitochondrial dna , biology , microbiology and biotechnology , sybr green i , real time polymerase chain reaction , mitochondrial disease , cytometry , lactic acidosis , mitochondrion , virology , viral load , biochemistry , antiretroviral therapy , human immunodeficiency virus (hiv) , gene
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. © 2012 International Clinical Cytometry Society