New molecular methods in prenatal diagnosis of aneuploidy mosaicism

非整倍体嵌合体产前诊断的新分子方法

• 批准号: 06670797
• 负责人: KANZAKI Susumu
• 金额: $ 1.34万
• 依托单位: Okayama University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06670797/
• 关键词: Prenatal diagnosis; Amniotic fluid cell; Chromosome abnormality; Mosaicism; FISH; PRINS; FISH法; 性染色体異常症; 多色FISH法; モガイシズム; 二色FISH法

The occurrence of mosaicism has been imposing a serious challenge to the prenatal diagnosis of chromosome abnormalities. The objective of this study is to investigate the frequency of chromosome mosaicism in prenatal diagnosis of amniotic fluid cells and to assess the utility of new molecular methods in the detection of mosaicism. The study subjects were 1,102 cases of amniotic fluid cell cultures. We categorized the mosaicism into a level 1 where a chromosome abnormality is seen in a single cell, level 2 where the abnormality is confined to plural cells in a single culture flask, and level 3 where the abnormality is seen in more than 2 flasks. A total of 77 (7.0%) cases were associated with any mosacism : 5 cases had the level 3 mosaicism [2 cases of trisomy 21, each case of trisomy 18, +inv dup (15) and t (14 ; 15)], 10 cases the level 2 mosaicism, and 62 cases the level 1 mosaicism. We applied the fluorescence in situ hybridization (FISH) and primed in situ hybridization (PRINS) methods to the diagnosis of the true mosaicism in the level 3 cases. Both FISH to alphoid DNA probes for chromosomes 15,18 and 21 and PRINS with primers specific to alphoid DNAs for chromosomes 18 and 21 showed significantly more signals on interphase nuclei in those cases than in normal controls. Furthermore, study in artificially made mosaic trisomy 18 showed that the FISH method can even detect a 12.5% of mosaicism. The very high frequency of chromosome mosaicism in amniotic fluid cell cultures suggest that much care should be taken to avoid misdiagnosis. We believe that FISH and PRINS methods which can analyze a great number of interphase cells in a short time are particularly useful in confirming the diagnosis of the true mosaicism.

嵌合体的出现给染色体异常的产前诊断带来了严峻的挑战。本研究的目的是调查羊水细胞产前诊断中染色体嵌合的频率，并评估新分子方法在嵌合检测中的效用。研究对象为1102例羊水细胞培养物。我们将镶嵌现象分为1级（在单个细胞中观察到染色体异常）、2级（异常仅限于单个培养瓶中的多个细胞）和3级（在2个以上培养瓶中观察到异常）。共有 77 例 (7.0%) 病例与任何嵌合体相关：5 例具有 3 级嵌合体 [2 例 21 三体性，各例 18 三体性，+inv dup (15) 和 t (14; 15)]，10 例具有 2 级嵌合体，62 例具有 1 级嵌合体。我们应用荧光原位杂交（FISH）和引物原位杂交（PRINS）方法来诊断3级病例中的真正嵌合体。 15,18 和 21 号染色体的 alphoid DNA 探针的 FISH 以及带有 18 和 21 号染色体的 alphoid DNA 特异性引物的 PRINS 在这些病例中均显示出间期核上的信号明显多于正常对照。此外，对人工制作的嵌合体18三体的研究表明，FISH方法甚至可以检测到12.5%的嵌合体。羊水细胞培养物中染色体嵌合的频率非常高，表明应格外小心以避免误诊。我们相信，FISH 和 PRINS 方法可以在短时间内分析大量间期细胞，对于确认真正嵌合体的诊断特别有用。

项目成果

Yokoyama Y et al.:"Cryptic pericentric inversion of chromosome 17 detected by fluorescence in situ hybridization in familial Miller Dieker syndrome" Amerlcan Journal of Medical Genetics. (in press). (1997)

Yokoyama Y 等人：“通过荧光原位杂交在家族性米勒迪克综合征中检测到 17 号染色体的隐性中心周倒位”美国医学遗传学杂志。

香山紀代子、楢原幸二、他: "羊水細胞の染色体分析におけるmosaicismの問題と対応" 日本小児科学会雑誌. 101・2. 196-196 (1997)

Kiyoko Kayama、Koji Narahara等人：“羊水细胞染色体分析中嵌合现象的问题和对策”日本儿科学会杂志101・2（1997）。

工藤尚文、平松祐司、横山裕司、辻一城他: "羊水細胞による出生前診断-モザイシズムを中心に-" 産婦人科の世界. 46. 41-45 (1994)

Naofumi Kudo、Yuji Hiramatsu、Yuji Yokoyama、Kazujo Tsuji 等人：“利用羊水细胞进行产前诊断 - 关注嵌合体 -”《妇产科世界》46. 41-45 (1994)。

Kiyoko Koyama, Koji Narahara, Yuji Yokoyama, Michito Teraoka et al.: "Mosaicism for chromosome abnormalities in amniotic fluid cell cultures-problem and solution" Nihonshonikagakkaizasshi. 101. 196 (1997)

Kiyoko Koyama、Koji Narahara、Yuji Yokoyama、Michito Teraoka 等：“羊水细胞培养中染色体异常的镶嵌现象 - 问题与解决方案”Nihonshonikagakkaizasshi。

松原恒則、横山裕司、楢原幸二、辻一城他: "FISH法の羊水細胞迅速診断への応用:異数性染色体異常とモザイシズムの同定に対する有用性と限界" 日本小児科学会雑誌. 99. 211- (1995)

Tsunenori Matsubara、Yuji Yokoyama、Koji Narahara、Kazujo Tsuji 等人：“FISH 方法在羊水细胞快速诊断中的应用：非整倍体染色体异常和嵌合体鉴定的有用性和局限性”，日本儿科学会杂志 99。 211-（1995）