Quantification of mitochondrial DNA using real-time polymerase chain reaction in patients with premature ovarian failure

Fertil Steril. 2005 Dec;84(6):1712-8. doi: 10.1016/j.fertnstert.2005.06.036.

Abstract

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.

Publication types

  • Clinical Trial
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Apoptosis
  • Feasibility Studies
  • Female
  • Genetic Testing / methods*
  • Humans
  • Mitochondria / genetics*
  • Primary Ovarian Insufficiency / genetics*
  • Primary Ovarian Insufficiency / pathology
  • Prospective Studies
  • Reverse Transcriptase Polymerase Chain Reaction / methods*