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A simple and rapid enzymatic assay for the branched-chain α-ketoacid dehydrogenase complex using high-performance liquid chromatography.

Authors :
Tajima, G.
Yofune, H.
Bahagia Febriani, A.
Nishimura, Y.
Ono, H.
Sakura, N.
Source :
Journal of Inherited Metabolic Disease; Sep2004, Vol. 27 Issue 5, p633-639, 7p
Publication Year :
2004

Abstract

Summary: Maple syrup urine disease (MSUD) is caused by a congenital defect of the branched-chain α-ketoacid dehydrogenase complex (BCKADC), and is one of the target disorders in newborn screening. However, it is not always easy to confirm the diagnosis; conventional methods of enzyme assay require cell culture, isolation of mitochondria, or radioisotope-labelled reagents, and disease-causing mutations can exist in any of the genes encoding the three enzyme subunits. To realize a practical test for diagnostic confirmation, we developed a simple and rapid enzymatic assay for BCKADC. In this procedure, the production of isovaleryl-CoA from 2-ketoisocaproic acid was measured using high-performance liquid chromatography. Detection of the BCKADC product was significantly reproducible depending on concentration of the substrates. We applied the assay to two patients with MSUD and demonstrated pathologically low levels of residual activity in both subjects. These results indicate that our method is a practical and sensitive assay for BCKADC, and that it can be a useful adjunct in newborn screening for MSUD. [ABSTRACT FROM AUTHOR]

Details

Language :
English
ISSN :
01418955
Volume :
27
Issue :
5
Database :
Complementary Index
Journal :
Journal of Inherited Metabolic Disease
Publication Type :
Academic Journal
Accession number :
51586126
Full Text :
https://doi.org/10.1023/B:BOLI.0000042988.31581.ed