Quantification of human mitochondrial DNA in a real time PCR

Wurmb‐Schwark, Nicole von; Higuchi, Russell; Fenech, Alan Paul; Elfstroem, C.; Meißner, Christoph; Oehmichen, M.; Cortopassi, Gino A

doi:10.1016/s0379-0738(02)00026-9

Cited by 60 publications

(30 citation statements)

References 19 publications

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“…If a particular deletion is known, the specificity and sensitivity of detection can be enhanced by designing a TaqMan oligonucleotide probe complementary to the specific junction sequence of the breakpoints. 18 We have used this approach to detect the common 4977-bp deletion and found that a heteroplasmic deletion as low as 0.01% can be detected (data not shown). The application of this method in clinical diagnosis is significant if different probes consistently show more than 10% deletion, an estimated reliable lower limit of detection sensitivity for clinical diagnosis.…”

Section: Discussionmentioning

confidence: 99%

“…Recently, real-time quantitative polymerase chain reaction (qPCR) method has been used to detect and quantify the common point mutations; 3243AϾG 14,15 and 8344AϾG 16,17 and the common 5-kb deletion. 18 More recently, this method was applied to the study of deletion, depletion, and overreplication. 13,19,20 In this study, we developed a comprehensive real-time qPCR method using various mtDNA and nuclear DNA probes to simultaneously detect and quantify mtDNA deletion, depletion, and over-replication in muscle specimens of patients with known deletions and in patients without identified mutations who showed mtDNA depletion and over-replication.…”

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confidence: 99%

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Simultaneous Detection and Quantification of Mitochondrial DNA Deletion(s), Depletion, and Over-Replication in Patients with Mitochondrial Disease

Bai

Wong²

2005

The Journal of Molecular Diagnostics

145

115

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

Simultaneous Detection and Quantification of Mitochondrial DNA Deletion(s), Depletion, and Over-Replication in Patients with Mitochondrial Disease

Bai

Wong²

2005

The Journal of Molecular Diagnostics

145

115

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“…External standards were prepared as described previously (von Wurmb-Schwark et al, 2002;May-Panloup et al, 2005) with minor modifications. PCR reactions were performed using iProof High Fidelity DNA Polymerase (Bio-Rad Laboratories) and 100 ng of genomic DNA.…”

Section: Downloaded Frommentioning

confidence: 99%

The Thiazolidinedione Pioglitazone Alters Mitochondrial Function in Human Neuron-Like Cells

Ghosh¹,

Patel²,

Rahn³

et al. 2007

Mol Pharmacol

101

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Thiazolidinediones alter cell energy metabolism. They are used to treat or are being considered for the treatment of disorders that feature mitochondrial impairment. Their mitochondrial effects, however, have not been comprehensively studied under long-term exposure conditions. We used the human neuronlike NT2 cell line to directly assess the long-term effects of a thiazolidinedione drug, pioglitazone, on mitochondria. At micromolar concentrations, pioglitazone increased mitochondrial DNA (mtDNA) content, levels of mtDNA and nuclear-encoded electron transport chain subunit proteins, increased oxygen consumption, and elevated complex I and complex IV V max activities. Pioglitazone treatment was also associated with increased cytoplasmic but reduced mitochondrial peroxide levels. Our data suggest that pioglitazone induces mitochondrial biogenesis and show that pioglitazone reduces mitochondrial oxidative stress in a neuron-like cell line. For these reasons pioglitazone may prove useful in the treatment of mitochondriopathies.

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“…In humans, the incidence of a 4,977-bp deletion in mtDNA known as the ''common deletion'' increases with age (10,11). Additionally, the occurrence of this deletion was four times greater in oral tissues of individuals who chewed areca nut, a major risk factor for oral and esophageal cancer, and the deletion was also induced in neuronal cells exposed to the mutagen ethidium bromide (12,13).…”

Section: Introductionmentioning

confidence: 99%

Arsenic Induced Mitochondrial DNA Damage and Altered Mitochondrial Oxidative Function: Implications for Genotoxic Mechanisms in Mammalian Cells

Partridge¹,

Huang

Hernandez-Rosa

et al. 2007

Cancer Research

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Arsenic is a well-established human carcinogen that is chronically consumed in drinking water by millions of people worldwide. Recent evidence has suggested that arsenic is a genotoxic carcinogen. Furthermore, we have shown that mitochondria mediate the mutagenic effects of arsenic in mammalian cells, as arsenic did not induce nuclear mutations in mitochondrial DNA (mtDNA)-depleted cells. Using the human-hamster hybrid A L cells, we show here that arsenic alters mitochondrial function by decreasing cytochrome c oxidase function and oxygen consumption but increasing citrate synthase function. These alterations correlated with depletion in mtDNA copy number and increase in large heteroplasmic mtDNA deletions. In addition, mtDNA isolated periodically from cultures treated continuously with arsenic did not consistently display the same deletion pattern, indicating that the mitochondrial genome was subjected to repeated and continuous damage. These data support the theory that the mitochondria, and particularly mtDNA, are important targets of the mutagenic effects of arsenic in mammalian cells. [Cancer Res 2007;67(11):5239-47]

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Quantification of human mitochondrial DNA in a real time PCR

Cited by 60 publications

References 19 publications

Simultaneous Detection and Quantification of Mitochondrial DNA Deletion(s), Depletion, and Over-Replication in Patients with Mitochondrial Disease

Simultaneous Detection and Quantification of Mitochondrial DNA Deletion(s), Depletion, and Over-Replication in Patients with Mitochondrial Disease

The Thiazolidinedione Pioglitazone Alters Mitochondrial Function in Human Neuron-Like Cells

Arsenic Induced Mitochondrial DNA Damage and Altered Mitochondrial Oxidative Function: Implications for Genotoxic Mechanisms in Mammalian Cells

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