Quantification of mitochondrial DNA using real-time polymerase chain reaction in patients with premature ovarian failure.
Cha KY, Lee SH, Chung HM, Baek KH, Cho SW, Kwack KB.
Department of Obstetrics and Gynecology, CHA General Hospital, Pochon CHA University, Seoul, Korea.
OBJECTIVE: To quantify mitochondrial DNA using real-time PCR in women with premature ovarian failure (POF) and a control group. DESIGN: Prospective study. SETTING: Genome Research Center for Reproductive Medicine and Infertility, Korea Ministry of Health & Welfare. PATIENT(S): Thrity patients with POF and 30 control individuals. INTERVENTION(S): The mitochondrial DNA content was quantified using real-time PCR. The effectiveness of the assay was determined by relative quantification using the comparative threshold cycle (CT) method. MAIN OUTCOME MEASURE(S): Relative quantification of mitochondrial DNA content. RESULT(S): The mitochondrial DNA content was significantly lower in the POF group than in the control group (0.58 +/- 0.38 vs. 1.15 +/- 0.67; P < .01). In both groups, there was a significant positive correlation between the mitochondrial DNA/28S rRNA ratio and mitochondrial DNA CT (control group: r = 0.774; P < .001; POF group: r = 0.556; P = .001) and a significant negative correlation between the mitochondrial DNA/28S rRNA ratio and 28S rRNA CT (control group: r = -0.677; P < .001; POF group: r = -0.627; P = .001). CONCLUSION(S): This study has established the clinical feasibility of quantifying amounts of mitochondrial DNA, relative to an internal standard, using real-time PCR. Further studies are warranted to elucidate the roles of apoptosis and mitochondrial function in the pathogenesis of POF.
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PMID: 16359970 [PubMed - indexed for MEDLINE]