title: 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.
abstract: Many of the previously described enzymatic assay methods for the diagnosis of medium-chain acyl-CoA dehydrogenase (MCAD) deficiency have been dependent upon the measurement of radioisotope-labeled co-products or reduction of electron acceptors. We have developed a direct assay 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 of 2-octenoyl-CoA was significantly reproducible. We applied the assay to samples from four infants suspected to have MCAD deficiency by tandem mass spectrometry (MS/MS) newborn screening conducted in the Hiroshima area of Japan. Three of them were proved to have pathologically reduced residual enzyme activities, although they were associated with various clinical and biochemical phenotypes. In addition, another symptomatic Japanese patient and her presymptomatic sibling who were detected by MS/MS selective screening were successfully diagnosed by our enzymatic assay. These results indicate that the method can be a useful confirmatory test for MS/MS screening of MCAD deficiency.
authors.affiliation: Department of Pediatrics, Hiroshima University Graduate School of Biomedical Sciences, Hiroshima 734-8551, Japan. ugoch@mac.com
authors.forename: Go
authors.initials: G
authors.lastname: Tajima
authors.forename: Nobuo
authors.initials: N
authors.lastname: Sakura
authors.forename: Hiroko
authors.initials: H
authors.lastname: Yofune
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Nishimura
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Ono
authors.forename: Yuki
authors.initials: Y
authors.lastname: Hasegawa
authors.forename: Ikue
authors.initials: I
authors.lastname: Hata
authors.forename: Masahiko
authors.initials: M
authors.lastname: Kimura
authors.forename: Seiji
authors.initials: S
authors.lastname: Yamaguchi
authors.forename: Yosuke
authors.initials: Y
authors.lastname: Shigematsu
authors.forename: Masao
authors.initials: M
authors.lastname: Kobayashi
mesh_terms: D000214:Acyl Coenzyme A / Q000032:analysis* / Q000378:metabolism; D042964:Acyl-CoA Dehydrogenase / Q000172:deficiency* / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D002675:Child, Preschool; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005260:Female; D005296:Ferrous Compounds / Q000378:metabolism; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D007564:Japan; D008214:Lymphocytes / Q000201:enzymology / Q000378:metabolism; D008297:Male; D008661:Metabolism, Inborn Errors / Q000175:diagnosis* / Q000201:enzymology / Q000235:genetics; D015997:Neonatal Screening / Q000379:methods / Q000592:standards; D015203:Reproducibility of Results; D017384:Sequence Deletion; D012689:Sequence Homology, Nucleic Acid; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods
chemical_list: D000214:Acyl Coenzyme A; D005296:Ferrous Compounds; C094936:octenoyl-coenzyme A; C064804:ferrocenium; C034681:octanoyl-coenzyme A; D042964:Acyl-CoA Dehydrogenase