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Korean Journal of Obstetrics & Gynecology 2004;47(1):16-24.
Published online January 1, 2004.
The Quantitative Analysis of Mitochondrial DNA Copy Number in Premature Ovarian Failure Patients Using the Real-time Polymerase Chain Reaction.
Jeong Hwan Kim, Sook Hwan Lee, Sung Won Cho, Hye Jin Jeong, Hyun Ah Kim, Yun Jung Lee, Joong Yul Na, In Sun Kim, Tae Ki Yoon, Jae Seong Kang
1Department of Obstetrics and Gynecology, Infertility Medical Center, CHA General Hospital, Seoul, Korea.
2Human Genetics Laboratory, Infertility Medical Center, CHA General Hospital, Seoul, Korea.
3Department of Obstetrics and Gynecology, College of Medicine, Korea University, Seoul, Korea.
4Department of Pathology, College of Medicine, Korea University, Seoul, Korea.
Abstract
OBJECTIVE
To design a screening method which identifies women with a potential of progression to the premature ovarian failure (POF) in near future, particularly among young women who have high FSH level but no symptoms and signs of POF. METHODS: Peripheral venous blood samples were collected from 30 patients with POF (age range, 19- 39 years) and from 30 controls with normal serum FSH level who had delivered two or more naturally conceived babies (age range, 32-39 years). mtDNA/28S rRNA ratio was determined by real-time quantitative PCR. Comparative threshold cycle (CT) method was adopted for the relative quantitation between two groups and the effectiveness of the method was evaluated. RESULTS: A significant decrease in mtDNA/28S rRNA ratio was found in the POF group (0.58 0.38) when compared with the control group (1.15 0.67) (P<0.01). In both control and POF groups, there was positive correlation between mtDNA and mtDNA/28S rRNA ratio (r=0.774, P<0.001; r=0.556, P=0.001, respectively) and negative correlation between 28S rRNA and mtDNA/28S rRNA ratio (r=-0.677, P<0.001; r=-0.627, P=0.001, respectively), which indicated the suitability of the method. CONCLUSION: This study showed a significant decrease in mtDNA copy number in the peripheral venous blood of POF patients. The comparative CT method was found to be an effective and efficient alternative for the screening purposes. With this basis, further studies on the early diagnostic and/or screening method for the POF might be enriched.
Key Words: Premature ovarian failure, Mitochondrial DNA, Real-time polymerase chain reaction
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