Development of a system for early diagnosis and molecular analysis of mitochondrial and peroxisomal fatty acid beta-oxidation defects in Japan.

我们研究了日本线粒体和过氧化物酶体脂肪酸β-氧化缺陷（FAOD）的早期诊断和分子分析系统的开发。结果如下：1）在尿有机酸的GC/MS分析中，采用稳定同位素稀释法，建立了酰基甘氨酸法检测线粒体FAOD的方法。2）建立了过氧化物酶体FAOD的血液滤纸筛选方法; 3）建立了线粒体FAOD的串联质谱酰基肉毒碱分析方法; 4）对日本确诊的线粒体FAOD和过氧化物酶体FAOD患者进行了调查。分别有40例和17例患者明确的线粒体和过氧化物酶体FAODs。5）建立了11例线粒体和10例过氧化物酶体FAODs的酶和免疫化学检测。6）在分子水平上研究了几例日本患者的新型线粒体FAODs，如极长链酰基辅酶A脱氢酶缺乏症或三功能蛋白缺乏症。7）一种新型过氧化物酶体酶，D-双功能蛋白，被发现，纯化和克隆。此外，还发现了一名酶缺乏症患者，并在酶和分子水平上进行了研究。8）基于本项目的结果，我们打算在日本开发一种该领域疾病的早期诊断和分子分析系统。它包括质谱、酶、免疫化学和遗传学方法。

We have studied on the development of a system for early diagnosis and molecular analysis of mitochondrial and peroxisomal fatty acid beta-oxidation defects (FAOD) in Japan. The results were as follows :1) In GC/MS analysis of urinary organic acids, acylglycine analysis for the detection of mitochondrial FAOD was established, using stable isotope dilution analysis. Furthermore, it was disclosed that the detection of dicarboxylepoxicides of long-chain carbon length was useful for diagnosis of peroxisomal FAOD.2) The screening method using blood filter paper was established for the peroxisomal FAOD.3) Acylcarnitnie analysis by tanden MS for the diagnosis of mitochondrial FAOD was established.4) Patients with mitochondrial and peroxisomal FAODs diagnosed in Japan were survelyed and investigated. There were 40 and 17 patients with definite mitochondrial and peroxisomal FAODs, respectively.5) Enzymatic and immunochemical detection for 11 mitochondrial and 10 peroxisomal FAODs was established.6) Several Japanese patients with novel mitochondrial FAODs, such as very-long chain acyl-CoA dehydrogenase deficiency or trifunctional protein deficiency, were investigated at the molecular levels.7) A novel peroxisomal enzyme, D-bifunctional protein, was discovered, purified and cloned. Furthermore, a patient with a deficiency of the enzyme was identified, and investigated at the enzymatic and molecular levels.8) Based on the results of this project, we are intending to develop a system of early diagnosis and molecular analysis for the disorders in this field in Japan. It includes mass spectrometric, enzymatic, immunochemical, and genetic approaches.