根据分子大小分离母体血浆中DNA序列检测β-地中海贫血父系遗传性胎儿点突变  

Detection of paternally inherited fetal point mutations for β-thalassemia using size-fractionated cell-free DNA in maternal plasma

作　　者：Di Naro E. Vitucci A. S.Hahn 张剑萍 

机构地区：[1]Laboratory for Prenatal Medicine, University Women' s Hospital, Department of Research, Spitalstrasse 21, CH 4031 Basel, Switzerland Dr

出　　处：《世界核心医学期刊文摘（妇产科学分册）》2005年第7期1-2,共2页Core Journal in Obstetrics/Gynecology

摘　　要：Context: Currently, fetal point mutations cannot be reliably analyzed from circulatory fetal DNA in maternal plasma, due to the predominance of maternal DNA sequences. However, analysis of circulatory fetal DNA sequences in maternal plasma have been shown to selectively enrich for fetal DNA molecules on the basis of a smaller molecular size than maternal DNA. Abstract Objective: To examine the prenatal analysis of 4 common β - thalassemia point mutations: IVSI- 1, IVSI- 6, IVSI- 110, and codon 39. Design, Setting, and Patients: A total of 32 maternal blood samples were collected at 10 to 12 weeks of gestation (mean, 10.7 weeks) between February 15, 2003, and February 25, 2004, in Bari, Italy, from women with risk for β - thalassemia in their newborns immediately prior to chorionic villous sampling. Samples in which the father and mother did not carry the same mutation were examined. Circulatory DNA was size- fractionated by gel electrophoresis and polymerase chain reaction (PCR) amplified with a peptide- nucleic- acid clamp, which suppresses amplification of the normal maternal allele. Presence of the paternal mutant allele was detected by allele- specific real- time PCR. Main Outcome Measure: Detection of paternally inherited β - globin gene point mutations. Results: Presence or absence of the paternal mutant allele was correctly determined in 6 (86% ) of 7 cases with the IVSI- 1 mutation, 4 (100% ) of 4 with the IVSI- 6 mutation, 5 (100% ) of 5 with the IVSI- 110 mutation, and 13 (81% ) of 16 with the codon 39 mutation. One false- positive test result was scored for the IVSI- 1 mutation. Two cases with the codon 39 mutation were classified as uncertain and 1 case was excluded due to lack of a diagnostic test result at the time of analysis. These results yielded an overall sensitivity of 100% and specificity of 93.8% , with classified cases removed. Conclusion: Our recently described technique of the size- fractionation of circulatory DNA in maternal plasma may be potentially useful for the noninvasive prenatContext: Currently, fetal point mutations cannot be reliably analyzed from circulatory fetal DNA in maternal plasma, due to the predominance of maternal DNA sequences. However, analysis of circulatory fetal DNA sequences in maternal plasma have been shown to selectively enrich for fetal DNA molecules on the basis of a smaller molecular size than maternal DNA. Abstract Objective: To examine the prenatal analysis of 4 common β - thalassemia point mutations: IVSI- 1, IVSI- 6, IVSI- 110, and codon 39. Design, Setting, and Patients: A total of 32 maternal blood samples were collected at 10 to 12 weeks of gestation (mean, 10.7 weeks) between February 15, 2003, and February 25, 2004, in Bari, Italy, from women with risk for β - thalassemia in their newborns immediately prior to chorionic villous sampling. Samples in which the father and mother did not carry the same mutation were examined. Circulatory DNA was size- fractionated by gel electrophoresis and polymerase chain reaction (PCR) amplified with a peptide- nucleic- acid clamp, which suppresses amplification of the normal maternal allele. Presence of the paternal mutant allele was detected by allele- specific real- time PCR. Main Outcome Measure: Detection of paternally inherited β - globin gene point mutations. Results: Presence or absence of the paternal mutant allele was correctly determined in 6 (86% ) of 7 cases with the IVSI- 1 mutation, 4 (100% ) of 4 with the IVSI- 6 mutation, 5 (100% ) of 5 with the IVSI- 110 mutation, and 13 (81% ) of 16 with the codon 39 mutation. One false- positive test result was scored for the IVSI- 1 mutation. Two cases with the codon 39 mutation were classified as uncertain and 1 case was excluded due to lack of a diagnostic test result at the time of analysis. These results yielded an overall sensitivity of 100% and specificity of 93.8% , with classified cases removed. Conclusion: Our recently described technique of the size- fractionation of circulatory DNA in maternal plasma may be potentially useful for the noninvasive prenat

关 键 词：DNA序列 Β-地中海贫血 父系遗传 点突变 分子大小 等位基因特异性 高危孕妇 Β-珠蛋白 选择性扩增 突变基因 

分 类 号：R714.5[医药卫生—妇产科学]