Clinical and Experimental Reproductive Medicine

The Korean Society for Reproductive Medicine (대한생식의학회)
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Reliability of the Single Cell PCR analysis for Preimplantation Genetic Diagnosis of Single Gene Disorders

단일 유전자 이상에 대한 착상전 유전진단을 위한 단일 세포 PCR 방법의 신뢰성

  • Choi, Hye Won (Laboratory of Reproductive Biology & Infertility, Samsung Cheil Hospital, Sungkyunkwan University School of Medicine) ;
  • Lee, Hyoung-Song (Laboratory of Reproductive Biology & Infertility, Samsung Cheil Hospital, Sungkyunkwan University School of Medicine) ;
  • Lim, Chun Kyu (Laboratory of Reproductive Biology & Infertility, Samsung Cheil Hospital, Sungkyunkwan University School of Medicine) ;
  • Koong, Mi Kyoung (Department of Obstetrics and Gynecology, Samsung Cheil Hospital, Sungkyunkwan University School of Medicine) ;
  • Kang, Inn Soo (Department of Obstetrics and Gynecology, Samsung Cheil Hospital, Sungkyunkwan University School of Medicine) ;
  • Jun, Jin Hyun (Laboratory of Reproductive Biology & Infertility, Samsung Cheil Hospital, Sungkyunkwan University School of Medicine)
  • 최혜원 (성균관대학교 의과대학 삼성제일병원 생식생물학 및 불임연구실) ;
  • 이형송 (성균관대학교 의과대학 삼성제일병원 생식생물학 및 불임연구실) ;
  • 임천규 (성균관대학교 의과대학 삼성제일병원 생식생물학 및 불임연구실) ;
  • 궁미경 (성균관대학교 의과대학 삼성제일병원 산부인과) ;
  • 강인수 (성균관대학교 의과대학 삼성제일병원 산부인과) ;
  • 전진현 (성균관대학교 의과대학 삼성제일병원 생식생물학 및 불임연구실)
  • Published : 2005.12.30
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Abstract

연구목적: 단일 유전자 이상에 대한 착상전 유전진단을 성공적으로 시행하기 위해서는 효과적이고 신뢰도가 높은 PCR 방법의 확립이 중요하다. 본 연구에서는 alkaline lysis와 duplex nested PCR 방법을 단일 림프구와 할구의 유전자 분석에 적용하여 그 효용성을 확인하고자 하였다. 재료 및 방법: 단일 유전자의 이상이 확인된 Duchenne muscular dystrophy (DMD), ornithine transcarbamylase (OTC) 결핍증과 epidermolysis bullosa (EB) 가계의 대상자들에서 채취한 단일 림프구와 공여 받은 배아의 할구를 이용하여 각각 PCR, restriction fragment length polymorphism (RFLP)와 direct DNA sequencing 분석을 시행하였다. 이러한 분석에서 유전자 증폭률 (amplification rate)과 두개의 allele 중에서 하나의 allele이 증폭되지 않는 allele drop-out (ADO) 빈도에 대해 살펴보았다. 결 과: 단일 림프구와 할구를 이용한 PCR 방법의 유전자 증폭률은 DMD에서 91.1%와 81.8%, OTC 결핍증에서 96.0%와 78.1%, EB에서 91.3%와 90.0%를 각각 나타냈으며, ADO 빈도는 OTC 결핍증에서 13.3%, EB에서 16.8%로 관찰되었다. 결 론: 본 연구에서 적용한 alkaline lysis와 duplex nested PCR 방법은 단일 유전자에 대한 착상전 유전진단에 성공적으로 적용할 수 있는 방법으로 생각되며, ADO 빈도를 최소화할 수 있는 효율적인 방법의 개발에 대한 지속적인 연구가 필요하다.

Keywords

References

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