title: Quantification of mitochondrial DNA using real-time polymerase chain reaction in patients with premature ovarian failure.
abstract: OBJECTIVE
  To quantify mitochondrial DNA using real-time PCR in women with premature ovarian failure (POF) and a control group.
  
  
  METHODS
  Prospective study.
  
  
  METHODS
  Genome Research Center for Reproductive Medicine and Infertility, Korea Ministry of Health & Welfare.
  
  
  METHODS
  Thrity patients with POF and 30 control individuals.
  
  
  METHODS
  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.
  
  
  METHODS
  Relative quantification of mitochondrial DNA content.
  
  
  RESULTS
  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).
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Obstetrics and Gynecology, CHA General Hospital, Pochon CHA University, Seoul, Korea.
authors.forename: Kwang-Yul
authors.initials: KY
authors.lastname: Cha
authors.forename: Sook-Hwan
authors.initials: SH
authors.lastname: Lee
authors.forename: Hyung-Min
authors.initials: HM
authors.lastname: Chung
authors.forename: Kwang-Hyun
authors.initials: KH
authors.lastname: Baek
authors.forename: Sung-Won
authors.initials: SW
authors.lastname: Cho
authors.forename: Kyu-Bum
authors.initials: KB
authors.lastname: Kwack
mesh_terms: D000328:Adult; D017209:Apoptosis; D005240:Feasibility Studies; D005260:Female; D005820:Genetic Testing / Q000379:methods*; D006801:Humans; D008928:Mitochondria / Q000235:genetics*; D016649:Primary Ovarian Insufficiency / Q000235:genetics* / Q000473:pathology; D011446:Prospective Studies; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*