title: Identification of a new mutation in medium-chain acyl-CoA dehydrogenase (MCAD) deficiency.
abstract: A mutation involving an A-to-G nucleotide replacement at position 985 of the medium-chain acyl-CoA dehydrogenase (MCAD) cDNA was found in homozygous form in 18 unrelated MCAD-deficient families and in heterozygous form in 4 families. By PCR amplification and sequencing of cDNA from a compound heterozygote, we have detected a new mutation in an MCAD-deficient patient in whom one MCAD allele produces mRNA that is missing 4 bp in the MCAD cDNA, while the other allele carries the A-to-G-985 mutation. The presence of this 4-bp deletion was confirmed in the patient's genomic DNA by dot-blot hybridization with allele-specific oligonucleotide probes and by restriction analysis of PCR products. A rapid screening test for this 4-bp deletion was developed, based on mismatched primer PCR amplification. The deletion created a new restrictive-enzyme site which yielded two DNA fragments. The 4-bp deletion was not found in the three remaining MCAD chromosomes not harboring the A-to-G-985 mutation, nor it was present in 20 chromosomes from 10 unrelated normal Caucasians. The PCR-based method for screening these two mutations can detect over 93% of all MCAD mutations.
authors.affiliation: Department of Pediatrics, Duke University Medical Center, Durham, NC 27710.
authors.forename: J H
authors.initials: JH
authors.lastname: Ding
authors.forename: B Z
authors.initials: BZ
authors.lastname: Yang
authors.forename: Y
authors.initials: Y
authors.lastname: Bao
authors.forename: C R
authors.initials: CR
authors.lastname: Roe
authors.forename: Y T
authors.initials: YT
authors.lastname: Chen
mesh_terms: D044944:Acyl-CoA Dehydrogenases / Q000172:deficiency / Q000235:genetics*; D000483:Alleles; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D006579:Heterozygote; D006720:Homozygote; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D010375:Pedigree; D016133:Polymerase Chain Reaction
chemical_list: D004247:DNA; D044944:Acyl-CoA Dehydrogenases