A duplex real-time PCR assay for detection of drug-induced mitochondrial DNA depletion in HepG2 cells.

Full Abstract

Screening of drug-induced mitochondrial DNA (mtDNA) depletion during early preclinical drug development is of major interest. Here we describe the establishment of a novel duplex calibrator-normalized real-time polymerase chain reaction (PCR) assay for rapid and reliable quantification of mtDNA in HepG2 cells. This assay involves quantification of an mtDNA target gene (cytochrome b) relative to a nuclear DNA (nDNA) reference gene (beta-actin) in one tube. The assay was evaluated for its precision, linearity, and reproducibility, and reliable detection of mtDNA depletion was demonstrated. Using this novel real-time PCR assay, drug-induced mtDNA depletion could be accurately detected.

Author information

Author/s: Höschele, Doris (D); Wiertz, Martina (M); Garcia Moreno, Inmaculada (I);

Affiliation: Federal Institute for Drugs and Medical Devices, D-53175 Bonn, Germany. hoeschel(-atsign-)bfarm.de

Journal and publication information

Publication Type: Journal Article

Journal: Analytical biochemistry (Anal Biochem), published in United States. (Language: eng)

Reference: 2008-Aug; vol 379 (issue 2) : pp 208-10

Dates: Created 2008/07/01; Completed 2008/10/09; Revised 2008/11/24;

PMID: 18541134, status: MEDLINE (last retrieval date: 2/18/2009, IMS Date: )

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