权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Development of non-invasive fetal DNA diagnosis from maternal blood and its clinical application

母血无创胎儿DNA诊断技术的发展及其临床应用

基本信息

批准号：
05807101
负责人：
TAKABAYASHI Haruo
金额：
$ 1.15万
依托单位：
Kanazawa Medical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1995
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05807101/
关键词：
fetal diagnosis DNA diagnosis maternal blood nucleated erythrocyte PCR FISH Percoll micromanipulation percoll法

项目摘要

Nucleated cells of fetal origin have been reported to be present in maternal blood in the majority of pregnancies. Fetal cells in maternal blood offer an alternative source of specimens to those obtained by invasive techniques such as amniocentesis, chorionicvillus sampling, or percutaneous umbilical blood sampling. The ability to retrieve fetal DNA infromation from maternal blood would enable all women to undergo fetal DNA analysis. The non-invasive recovery of fetal cells from maternal blood has great potential to revolutionize fetal medicine. Several attempts have been made to detect and retrieve fetal nucleated cells including nucleated erythrocytes (NRBCs), leukocytes and trophoblasts in maternal blood. Because the number of fetal nucleated cells in maternal blood is extremely limited, there have been some trials using flow sorting or magnetic sorting as the collection method. NRBCs are a good target cell population for fetal DNA diagnosis because they are unlikely to circulate in the peripheral blood of a normal adult. The data indicated that NRBCs were detected after more than 5 weeks of gestation. NoNRBCs were found in the adult controls. These fetal NRBCs were successfully retrieved using a micromanipulator under a microscope. It was revealed that the single NRBCs removed from maternal blood were fetal in origin, because a Y-specific band could be detected in DNA from a single male cell after PCR amplification. We were also successful in applying the FISH technique to the May-Giemsa-stained NRBCs of fetuses. This new technique opens up fetal DNA diagnosis from maternal blood during the first trimester of pregnancy to the whole population because there is no risk to the fetus or the mother.

有报道称，大多数孕妇的母体血液中都存在胎儿来源的有核细胞。孕妇血液中的胎儿细胞提供了一种替代侵入性技术(如羊膜穿刺术、绒毛取样或经皮脐血取样)获得的样本来源。从母体血液中提取胎儿DNA信息的能力将使所有妇女都能接受胎儿DNA分析。从母血中无创回收胎儿细胞具有极大的潜力，将给胎儿医学带来革命性的变化。人们曾多次尝试检测和提取孕妇血液中的胎儿有核细胞，包括有核红细胞(NRBCs)、白细胞和滋养细胞。由于母血中胎儿有核细胞的数量极其有限，因此已经有一些尝试使用流动分选或磁分选作为采集方法。NRBC是胎儿DNA诊断的良好靶细胞群，因为它们不太可能在正常成年人的外周血液中循环。结果表明，NRBC在妊娠5周以上即可检出。成人对照组未发现NRBC。这些胎儿的NRBCs是在显微镜下使用显微操作器成功提取的。结果表明，从母血中提取的单个NRBCs来自胎儿，因为在单个男性细胞的DNA中可以检测到一条Y-特异性条带。我们还成功地将FISH技术应用于5月-Giemsa染色的胎儿NRBC。这项新技术使怀孕前三个月孕妇血液中的胎儿DNA诊断面向全体人群，因为这对胎儿或母亲没有风险。