Enzymatic diagnosis of medium-chain acyl-CoA dehydrogenase deficiency by detecting 2-octenoyl-CoA production using high-performance liquid chromatography: a practical confirmatory test for tandem mass spectrometry newborn screening in Japan.

摘要: Many of the previously described enzymatic assay methods for diagnosis medium-chain acyl-CoA dehydrogenase (MCAD) deficiency have been dependent upon measurement radioisotope-labeled co-products or reduction electron acceptors. We developed a direct method to detect 2-enoyl-CoA production using high-performance liquid chromatography (HPLC). Crude cell lysate prepared from lymphocytes were incubated with n-octanoyl-CoA and ferrocenium hexafluorophosphate. The detection 2-octenoyl-CoA was significantly reproducible. applied samples four infants suspected MCAD by tandem mass spectrometry (MS/MS) newborn screening conducted in Hiroshima area Japan. Three them proved pathologically reduced residual enzyme activities, although they associated various clinical biochemical phenotypes. In addition, another symptomatic Japanese patient her presymptomatic sibling who detected MS/MS selective successfully diagnosed our assay. These results indicate that can be useful confirmatory test deficiency.