Quantification of mitochondrial DNA using real-time polymerase chain reaction in patients with premature ovarian failure.
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.
Cha
Kwang-Yul
KY
Department of Obstetrics and Gynecology, CHA General Hospital, Pochon CHA University, Seoul, Korea.
Lee
Sook-Hwan
SH
Chung
Hyung-Min
HM
Baek
Kwang-Hyun
KH
Cho
Sung-Won
SW
Kwack
Kyu-Bum
KB
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*