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Fetal diagnosis using fetal erythroid progenitor cells in maternal blood

利用母血中的胎儿红系祖细胞进行胎儿诊断

基本信息

批准号：
10671546
负责人：
MIHARU Norio
金额：
$ 0.96万
依托单位：
HIROSHIMA UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 2000
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10671546/
关键词：
PCR DNA amniocentesis serum fetus 胎児赤血球 fluoressence in situ hybridization FISH

项目摘要

Peripheral blood samples were obtained from 61 pregnant women at 10-17 weeks of gestation before amniocentesis. DNA was extracted from 800 microL of each plasma or serum sample. To detect the Y-chromosome-specific sequences DYS14 and DYZ3 in the maternal plasma and serum, 40 cycles of PCR were carried out for each DNA extract. The PCR products were analyzed by 2.5% agarose gel electrophoresis and ethidium bromide staining, and the results were compared with the results of the cytogenetic analyses of amniocentesis.Cytogenetic analysis of amniocentesis revealed that 31 pregnant women had a male fetus and the remaining 30 pregnant women had a female fetus. Both DYS14 and DYZ3 were detected in 27 of the 31 plasma samples obtained from pregnant women carrying a male fetus and in all of 31 serum samples obtained from the same women. Neither DYS14 nor DYZ3 was detected in either the plasma or serum samples obtained from any of the 30 pregnant women carrying a female fetus.PCR analysis of mate … More rnal serum can be used to diagnose fetal gender.Serum samples were obtained from pregnant women at gestational ages ranging from 15 to 17 weeks, prior to their undergoing amniocentesis. In total, we examined 70 samples consisting of 55 cases of pregnancy with 46, XY, 5 cases with 47, XY, +21, 3 cases with 47, XY, +18, a single case with 46, XY, dup (1) and 2 cases with twins of 46, XY, and 4 cases with 46, XX which were used as negative controls. We measured the concentration of the SRY sequence as a molecular marker for fetal DNA using real-time quantitative polymerase chain reaction (PCR) assay.The SRY sequence was detectable and measurable when the fetuses were male except for one case with 47, XY, +18. This case showed fetal growth retardation and bradycardia. No amplification signals of the SRY sequence were detected when the fetuses were female. The mean concentration of fetal DNA in maternal serum was 31.5 copies/ml in the pregnancy with 46, XY, 23.5 copies/ml in the pregnancies with 47, XY, +21 and 21.5 copies/ml in the pregnancies with 46, XY, +18. There were no significant differences in the concentration of fatal DNA between pregnancies with fetuses of normal karyotype and those with fetuses of abnormal karyotype. Less

在羊膜穿刺术前采集61例妊娠10-17周的孕妇外周血样本。从每个血浆或血清样品中提取800微升DNA。为了检测母体血浆和血清中y染色体特异性序列DYS14和DYZ3，每种DNA提取液进行40个PCR循环。PCR产物经2.5%琼脂糖凝胶电泳和溴化乙啶染色分析，并与羊膜穿刺术细胞遗传学分析结果进行比较。羊膜穿刺术的细胞遗传学分析显示，31名孕妇有男性胎儿，其余30名孕妇有女性胎儿。从携带男性胎儿的孕妇获得的31份血浆样本中有27份检测到DYS14和DYZ3，从同一名妇女获得的所有31份血清样本中均检测到DYS14和DYZ3。在30名怀孕女性的血浆和血清样本中均未检测到DYS14和DYZ3。更多的血清可用于胎儿性别的诊断。血清样本来自孕龄15至17周的孕妇，在进行羊膜穿刺术之前。我们共检查了70例样本，其中55例妊娠46、XY， 5例47、XY、+21,3例47、XY、+18,1例46、XY、dup(1)， 2例双胞胎46、XY， 4例46、XX作为阴性对照。我们使用实时定量聚合酶链反应（PCR）法测定SRY序列的浓度，作为胎儿DNA的分子标记。除1例47、XY、+18外，所有男性胎儿均可检测到SRY序列。本病例表现为胎儿生长迟缓和心动过缓。当胎儿为雌性时，未检测到SRY序列的扩增信号。46、XY孕妇血清中胎儿DNA平均浓度为31.5拷贝/ml， 47、XY、+21孕妇血清中胎儿DNA平均浓度为23.5拷贝/ml， 46、XY、+18孕妇血清中胎儿DNA平均浓度为21.5拷贝/ml。核型正常胎和核型异常胎的致死性DNA浓度差异无统计学意义。少