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대한생식의학회지 제32권 제1호 2010년
근이영양증에 대한 착상전 유전진단에서 Duplex-nested PCR과 Fluorescent PCR 방법의 효용성
성균관대학교 의과대학 삼성제일병원 생식생물학 및 불임연구실1, 유전학연구실2, 산부인과학교실3
이형송1, 최혜원1, 임천규1, 박소연2, 김진영3, 궁미경3, 전진현1*, 강인수3,
Efficacy of Duplex-nested PCR and Fluorescent PCR in the Preimplantation Genetic Diagnosis for Duchenne Muscular Dystrophy
Hyoung-Song Lee1, Hye Won Choi1, Chun Kyu Lim1, So Yeon Park2, Jin Young Kim3, Mi Kyoung Koong3, Jin Hyun Jun1*, Inn Soo Kang3
1Laboratory of Reproductive Biology and Infertility, 2Laboratory of Medical Genetics, 3Department of Obstetrics and Gynecology, Samsung Cheil Hospital and Women's Healthcare Center, Sungkyunkwan University School of Medicine
Objective: Preimplantation genetic diagnosis (PGD) is reserved for couples with a risk of transmitting a serious and incurable disease, and hence avoids the undesirable therapeutic abortion. In this study, we evaluated the efficacy of PGD for Duchenne muscular dystrophy (DMD) cases by the fluorescent PCR with polymorphic linked markers and the conventional duplex-nested PCR methods. Methods: Biopsy of one or two blastomeres was done from the embryos fertilized by ICSI on the third day after fertilization. We performed two cases of PGD-DMD by the duplex-nested PCR for the causative mutation loci and the SRY gene on Y chromosome. The triplex fluorescent PCR for the mutation loci, the SRY gene and the polymorphic microsatellite marker on X chromosome was applied for two cases of PGD-DMD. Results: By the duplex-nested PCR, successful diagnosis rate was 95.5% (21/22), but we could not discriminate the female embryos whether normal or carrier in this X-linked recessive disease. However, the triplex fluorescent PCR method showed 100% (27/27) of successful diagnosis rate, and all female embryos (n=17) were distinguished normal (n=10) from carrier (n=7) embryos. Unaffected and normal embryos were transferred into mother's uterus after diagnosis. A healthy normal male was achieved after PGD with the duplex-nested PCR method and a twin, a male and a female, were delivered with triplex fluorescent PCR method. The normality of dystrophin gene was confirmed by amniocentesis and postnatal genetic analysis in all offsprings. Conclusion: The fluorescent PCR with polymorphic marker might be useful in improving the specificity and reliability of PGD for single gene disorders.
키워드 : Preimplantation genetic diagnosis (PGD), Duchenne muscular dystrophy (DMD), Duplexnested PCR, Fluorescent PCR, Polymorphic marker
교신저자 : hslee99@unitel.co.kr
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