Journal of Genetic Medicine

Korean Society of Medical Genetics and Genomics (대한의학유전학회)
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Effective Method for Extraction of Cell-Free DNA from Maternal Plasma for Non-Invasive First-Trimester Fetal Gender Determination: A Preliminary Study

  • Lim, Ji-Hyae (Laboratory of Medical Genetics, Medical Research Institute, Cheil General Hospital and Women's Healthcare Center) ;
  • Park, So-Yeon (Laboratory of Medical Genetics, Medical Research Institute, Cheil General Hospital and Women's Healthcare Center) ;
  • Kim, Shin-Young (Laboratory of Medical Genetics, Medical Research Institute, Cheil General Hospital and Women's Healthcare Center) ;
  • Kim, Do-Jin (Laboratory of Medical Genetics, Medical Research Institute, Cheil General Hospital and Women's Healthcare Center) ;
  • Kim, Mee-Jin (Laboratory of Medical Genetics, Medical Research Institute, Cheil General Hospital and Women's Healthcare Center) ;
  • Yang, Jae-Hyug (Department of Obstetrics and Gynecology, Cheil General Hospital and Women's Healthcare Center KwanDong University college of Medicine) ;
  • Kim, Moon-Young (Department of Obstetrics and Gynecology, Cheil General Hospital and Women's Healthcare Center KwanDong University college of Medicine) ;
  • Kim, Min-Hyoung (Department of Obstetrics and Gynecology, Cheil General Hospital and Women's Healthcare Center KwanDong University college of Medicine) ;
  • Han, Ho-Won (Department of Obstetrics and Gynecology, Cheil General Hospital and Women's Healthcare Center KwanDong University college of Medicine) ;
  • Choi, Kyu-Hong (Department of Obstetrics and Gynecology, Cheil General Hospital and Women's Healthcare Center KwanDong University college of Medicine) ;
  • Ryu, Hyun-Mee (Laboratory of Medical Genetics, Medical Research Institute, Cheil General Hospital and Women's Healthcare Center)
  • Received : 2010.05.27
  • Accepted : 2010.06.18
  • Published : 2010.03.01
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Abstract

Purpose: To find the most effective method for extraction of cell-free DNA (cf-DNA) from maternal plasma, we compared a blood DNA extraction system (blood kit) and a viral DNA extraction system (viral kit) for non-invasive first-trimester fetal gender determination. Materials and Methods: A prospective cohort study was conducted with maternal plasma collected from 44 women in the first-trimester of pregnancy. The cf-DNA was extracted from maternal plasma using a blood kit and a viral kit. Quantitative fluorescent-polymerase chain reaction (QF-PCR) was used to detect the SRY gene and AMEL gene. The diagnostic accuracy of the QF-PCR results was determined based on comparison with the final delivery records. Results: A total of 44 women were tested, but the final delivery record was only obtained in 36 cases which included 16 male-bearing and 20 female-bearing pregnancies. For the blood kit and viral kit, the diagnostic accuracies for fetal gender determination were 63.9% (23/36) and 97.2% (35/36), respectively. Conclusion: In non-invasive first-trimester fetal gender determination by QF-PCR, using a viral kit for extraction of cf-DNA may result in a higher diagnostic accuracy.

목 적: 모체 혈장으로부터 가장 효과적으로 세포 유리 DNA(cell free DNA, cf-DNA)를 추출하는 방법을 찾기 위해 우리는 viral DNA 추출 방법과 일반 혈액DNA 추출 방법을 이용하여 비침습적 임신 초기 태아 성별 확인 결과를 비교하였다. 대상 및 방법: 임신 초기 44명의 임산부로부터 모여진 모체 혈장을 통한 전향적 연구가 구성되었다. Cf-DNA는 viral DNA 추출 방법과 일반 혈액 DNA 추출 방법을 이용하여 각각 추출되었다. 정량 형광-중합효소 연쇄 반응(QF-PCR)을 이용하여SRY 와AMXY 유전자를 검출하였다. QF-PCR의 진단 정확도는 최종 분만 기록을 토대로 결정하였다. 결 과: 전체 44명의 여성이 실험에 참여하였지만, 최종 분만 기록은 단지 36명의 여성에서 획득하였다. 이들 중 16명은 남아를 20명은 여아를 임신하였다. 두 추출 방법에서 태아 성별의 진단적 정확도는 일반 혈액 DNA 추출 방법에 경우 63.9% (23/26)였으며 viral DNA 추출 방법에 경우 97.2% (35/36) 였다. 결 론: QF-PCR을 이용한 비침습적 임신초기 태아 성별 확인에 있어 viral kit를 사용하는 것이 높은 진단적 정확도를 이끌 수 있을 것으로 사료된다.

Keywords

References

  1. Lo YM, Corbetta N, Chamberlain PF, Rai V, Sargent IL, Redman CW, et al. Presence of fetal DNA in maternal plasma and serum. Lancet 1997;350:485-7. https://doi.org/10.1016/S0140-6736(97)02174-0
  2. Lo YM, Hjelm NM, Fidler C, Sargent IL, Murphy MF, Chamberlain PF, et al. Prenatal diagnosis of fetal RhD status by molecular analysis of maternal plasma. N Engl J Med 1998;339:1734-38. https://doi.org/10.1056/NEJM199812103392402
  3. Rijnders RJ, van der Schoot CE, Bossers B, de Vroede MA, Christiaens GC. Fetal sex determination from maternal plasma in pregnancies at risk for congenital adrenal hyperplasia. Obstet Gynecol 2001;98:374-8. https://doi.org/10.1016/S0029-7844(01)01480-6
  4. Bianchi DW, Avent ND, Costa JM, van der Schoot CE. Noninvasive prenatal diagnosis of fetal Rhesus D: ready for Prime(r) Time. Obstet Gynecol 2005;106: 841-4. https://doi.org/10.1097/01.AOG.0000179477.59385.93
  5. Geifman-Holtzman O, Grotegut CA, Gaughan JP. Diagnostic accuracy of noninvasive fetal Rh genotyping from maternal blood: a meta-analysis. Am J Obstet Gynecol 2006;195:1163-73. https://doi.org/10.1016/j.ajog.2006.07.033
  6. Scheffer PG, van der Schoot CE, Page-Christiaens GC, Bossers B, van Erp F, de Haas M. Reliability of fetal gender determination using maternal plasma. Obstet Gynecol 2010;115:117-26. https://doi.org/10.1097/AOG.0b013e3181c3c938
  7. Chan KC, Zhang J, Hui AB, Wong N, Lau TK, Leung TN, et al. Size distributions of maternal and fetal DNA in maternal plasma. Clin Chem 2004;50:88-92. https://doi.org/10.1373/clinchem.2003.024893
  8. Li Y, Zimmermann B, Rusterholz C, Kang A, Holzgreve W, Hahn S. Size separation of circulatory DNA in maternal plasma permits ready detection of fetal DNA polymorphisms. Clin Chem 2004;50:1002-11. https://doi.org/10.1373/clinchem.2003.029835
  9. Rijnders RJP, Van Der Luijt RB, Peters ED, Goeree JK, Van Der Schoot CE, Ploos Van Amstel JK, et al. Earliest gestational age for fetal gendering in cell-free maternal plasma. Prenat Diagn 2003;23:1042-4. https://doi.org/10.1002/pd.750
  10. Clausen FB, Krog GR, Rieneck K, Dziegiel MH. Improvement in fetal DNA extraction from maternal plasma. Evaluation of the NucliSens Magnetic Extraction system and the QIAamp DSP Virus Kit in comparison with the QIAamp DNA Blood Mini Kit. Prenat Diagn 2007; 27:6-10. https://doi.org/10.1002/pd.1605
  11. Legler TJ, Liu Z, Mavrou A, Hromadnikova I, Galbiati S, Meaney C, et al. Workshop report on the extraction of foetal DNA from maternal plasma. Prenat Diagn 2007;27:824-829. https://doi.org/10.1002/pd.1783
  12. Gonzalez-Gonzalez C, Garcia-Hoyos M, Trujillo-Tiebas MJ, Lorda-Sanchez I, de Alba MR, Infantes F, et al. Application of fetal DNA detection in maternal plasma: a prenatal diagnosis unit experience. J Histochem Cytochem 2005;53:307-14. https://doi.org/10.1369/jhc.4A6400.2005
  13. Pertl B, Weitgasser U, Kopp S, Kroisel PM, Sherlock J, Adinolfi M. Rapid detection of trisomies 21 and 18 and gendering by quantitative fluorescent multiplex PCR. Hum Genet 1996;98:55-9. https://doi.org/10.1007/s004390050159
  14. Pertl B, Kopp S, Kroisel PM, Tului L, Brambati B, Adinolfi M. Rapid detection of chromosome aneuploidies by quantitative fluorescence PCR: First application on 247 chorionic villus samples. J Med Genet 1999;36:300-3.
  15. Schmidt W, Jenderny J, Hecher K, Hackelo er BJ, Kerber S, Kochhan L, et al. Detection of aneuploidy in chromosomes X, Y, 13, 18 and 21 by QF-PCR in 662 selected pregnancies at risk. Mol Hum Reprod 2000;6:855-60. https://doi.org/10.1093/molehr/6.9.855
  16. Cirigliano V, Ejarque M, Canadas MP, Lloveras E, Plaja A, Perez MM, et al. Clinical application of multiplex quantitative fluorescent polymerase chain reaction (QF-PCR) for the rapid prenatal detection of common chromosome aneuploidies. Mol Hum Reprod 2001;7:1001-6. https://doi.org/10.1093/molehr/7.10.1001