DEVELOPMENT OF MASS SCREENING METHOD FOR CONGENITAL BILIARY ATRESIA.

先天性胆道闭锁的大规模筛查方法的开发。

• 批准号: 07557297
• 负责人: TOHMA Masahiko
• 金额: $ 0.83万
• 依托单位: HEALTH SCIENCES UNIVERSITY OF HOKKAIDO
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07557297/
• 关键词: FETAL BILE ACID; CONGENITAL BILIARY ATRESIA; NEONATAL LIVER DISORDER; GAS CHROMATOGRAPHY-MASS SPECTROMETRY; HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY; ENZYME IMMUNOASSAY; MASS SCREENING; DRIED BLOOD SPOTTED FILER PAPER

The estsablishment of the diagnosis of congenital liver disorder is urgently required for the remedy of liver disorder in early life. The metabolism of bile acids in fetus and neonate is thought to be different from that in adults. The aim of this reaserch is the developement of quantitative analysis of fetal bile acids by GC-MS and immunological assay, elucidation of the profiles of bile acids with hepatobiliary diseases, and establishment of the precise diagnosis and mass screening of congenital biliary atresia.1. A highly sensitive GC-MS determination of the fetal bile acids in neonatal serum and dried blood spot disc has been developed by capillary GC-MS using selected ion monitoring with negative ion chemical ionization. Diagnostic bile acid analysis of neonatal liver diseases was accomplished by GC-MS.2. An improved quantitative analysis of the conjugated fetal bile acids in urine of newborns has been developed by HPLC employing labeling with 1-anthroyl cyanide and fluorescence detection.3. A highly sensitive enzyme immunoassay of conjugated 1beta-hydroxycholic acid in heterologous combination has been developed using horse radish peroxidase labeled antigen. Concentrations of the conjugated bile acid in dried blood spotted onto filter paper in newborns with congenital biliary atresia were significantly higher than those of normal newborns, showing the possibility of mass screening for the disorder.

建立先天性肝病的诊断是早期治疗肝病的迫切需要。胎儿和新生儿胆汁酸的代谢被认为与成人不同。本研究旨在发展气相色谱-质谱联用及免疫检测胎儿胆汁酸的定量分析，阐明胆汁酸与肝胆疾病的关系，建立先天性胆道闭锁的精确诊断和大规模筛查方法。建立了采用负离子化学电离选择性离子监测的毛细管气相色谱-质谱法测定新生儿血清和干血斑盘中胎儿胆汁酸的方法。采用气相色谱-质谱法对新生儿肝脏疾病进行胆汁酸诊断分析。采用高效液相色谱法（HPLC）对新生儿尿液中共轭胎儿胆汁酸进行了改进的定量分析。采用马萝卜过氧化物酶标记抗原，建立了一种高灵敏度的异种结合β -羟基胆酸的酶免疫测定方法。先天性胆道闭锁新生儿滤纸上干血中共轭胆汁酸浓度明显高于正常新生儿，提示有可能对该疾病进行大规模筛查。

项目成果

Masahiko Tohma: "Determination of Fetal Bile Acids and Related Steroidal Compounds and Their Profile in Neonatal Biological Fluids." YAKUGAKU ZASSHI. 116 (10). 753-775 (1996)

Masahiko Tohma：“胎儿胆汁酸和相关类固醇化合物的测定及其在新生儿生物液中的分布。”

Teruki Yoshimura: "High-Performance Liquid Chromatographic Determination of conjugated and Unconjugated 3-Oxo-Δ^4- and 3-Oxo-Δ^<4,6>-Bile Acids in Human Urine." J.Liq.Chrom.& Rel.Technol. 20(3). 923-934 (1997)

Teruki Yoshimura：“高效液相色谱测定人尿液中缀合和非缀合的 3-Oxo-Δ^4- 和 3-Oxo-Δ^<4,6>-胆汁酸。J.Liq.Chrom.& Rel。” .技术20(3)。

Takao Kurosawa: "Synthsis of 19-Hydroxylated Bile Acids and Identification of 3α,7α,12α,19-Tetrahydroxy-5β-cholan-24-oic Acid in Human Neonatal Urine." Chem.Pharm.Bull.43. 1551-1557 (1995)

Takao Kurosawa：“人类新生儿尿液中 19-羟基胆汁酸的合成和 3α,7α,12α,19-四羟基-5β-cholan-24-oic 酸的鉴定。”1551-1557（ 1995)

吉村昭毅: "胎児性胆汁酸測定における3α-ヒドロキシステロイドデヒドゲナーゼの特異性." 分析化学. 44. 865-869 (1995)

Akiyoshi Yoshimura：“胎儿胆汁酸测量中 3α-羟基类固醇脱氢酶的特异性。” 44. 865-869 (1995)

Ryoichi Yamakawa: "Urinary 1β-Hydroxyursodeoxycholic Acid during Ursodeoxycholic Acid Therapy." J.Gastroenterol.31. 917-918 (1996)

Ryoichi Yamakawa：“熊去氧胆酸治疗期间的尿液 1β-羟基熊去氧胆酸。”J.Gastroenterol.31 (1996)。