Urease-pretreatment, stable isotope dilution and gas chromatographic mass spectrometrie application for the chemical diagnosis of inborn errors of metabolism

脲酶预处理、稳定同位素稀释和气相色谱质谱在先天性代谢缺陷化学诊断中的应用

• 批准号: 11672312
• 负责人: KUHARA Tomiko
• 金额: $ 1.98万
• 依托单位: Kanazawa Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11672312/
• 关键词: GC; MS; Inborn error of metabolism; chemical diagnosis; pyrimidine degradation; homocystinuria; Pilot study of neonatal screening; folate deficiency; Propionic acidemia; 先天性代謝疾患; 自動前処理; 診断システム; 新生児スクリーニング; 5-フルオロウラシル; ろ紙尿; 代謝性疾患; 新生児マススクリー

1. In pyrimidine degradation three enzymes, PDH, DHP and UPase, are involved. In these enzyme deficiecies, pyrimidine analogues used for cancer chemotherapy cause serious side-effects. To prevent such severe side-effects, we developed a method by which specific, rapid and sensitive chemical diagnosis can made In "Modern Drug Discovery" published by American Chemical Society evaluated our method in "News in Brief".2. Using urease-pretreatment, stable isotope dilution and GC/MS, we established a method to make differential chemical diagnosis of homocystinuria. We also confirmed that this method also detect folate deficiency secondarily caused by homocystinuria type I.3. In the pilot study of neonatal screening, propionic acidemia )PCCD) and α-ketoadipic aciduria were found to occure 1 to 10,000〜20,000 babies. To detect mild type PCCD was supposed to be rather difficult in MS/MS using blood spots (J. Inher. Metab. Dis., (in press)).

1.在嘧啶降解中涉及三种酶，PDH、DHP和UPase。在这些酶缺乏的情况下，用于癌症化疗的嘧啶类似物会引起严重的副作用。为了避免这些严重的副作用，我们开发了一种特异、快速、灵敏的化学诊断方法。美国化学学会出版的“现代药物发现”杂志在“新闻简报”中对我们的方法进行了评价。采用尿素酶预处理、稳定同位素稀释和GC/MS技术，建立了同型胱氨酸尿症的鉴别诊断方法。我们还证实，这种方法也检测叶酸缺乏症继发引起的同型胱氨酸尿症I型。在新生儿筛查的初步研究中，发现丙酸血症（PCCD）和α-酮己二酸尿症发生在1至10，000至20，000名婴儿中。在MS/MS中使用血斑检测轻度型PCCD被认为是相当困难的（J. Inher. Metab. Dis.，(in press）。

项目成果

Kuhara,T.: "Gas chromatographic-mass spectrometric newborn screening and diagnosis of propionic acidemia by targeting methylcitrate in dried filter-paper urine samples"J. Inher. Metab. Dis.. (in press). (2002)

Kuhara，T.：“通过针对干燥滤纸尿液样本中的柠檬酸甲酯进行气相色谱-质谱法新生儿丙酸血症筛查和诊断”J。

T.Kuhara: "Differential Diagnosis of Homocystinuria by Urease-treatment, isotope-dilution and gas chromatography-mass spectrometry"J.Chromatogr.B. 742. 59-70 (2000)

T.Kuhara：“通过脲酶处理、同位素稀释和气相色谱-质谱法对同型半胱氨酸尿症进行鉴别诊断”J.Chromatogr.B.

Kuhara,T.: "Differential Diagnosis of Homocystinuria by Urease-treatment, isotope-dilution and gas chromatography-mass spectrometry"J. Chromatogr. B.. 742. 59-70 (2000)

Kuhara，T.：“通过脲酶处理、同位素稀释和气相色谱-质谱法对同型半胱氨酸尿症进行鉴别诊断”J。

久原とみ子: "有機酸の代謝異常"臨床検査. 44・1. 57-67 (2000)

久原富子：“有机酸代谢紊乱”临床检查44・1（2000）。

H.Peng: "Asymptomatic α-ketoadipic aciduria detected during a pilot study of neonatal urine screening"Acta Paediatr.. 88. 911-914 (1999)

H.Peng：“新生儿尿液筛查试验中检测到的无症状 α-酮己二酸尿症”Acta Paediatr.. 88. 911-914 (1999)