title: Application of multiplex ligation-dependent probe amplification, and identification of a heterozygous Alu-associated deletion and a uniparental disomy of chromosome 1 in two patients with 3-hydroxy-3-methylglutaryl-CoA lyase deficiency.
abstract: Mitochondrial 3-hydroxy-3-methylglutaryl-CoA lyase (HMGCL) deficiency is an autosomal recessive disorder affecting the leucine catabolic pathway and ketone body synthesis, and is clinically characterized by metabolic crises with hypoketotic hypoglycemia, metabolic acidosis and hyperammonemia. In the present study, we initially used PCR with genomic followed by direct sequencing to investigate the molecular genetic basis of HMGCL deficiency in two patients clinically diagnosed with the condition. Although we identified a mutation in each patient, the inheritance patterns of these mutations were not consistent with disease causation. Therefore, we investigated HMGCL using multiplex ligation-dependent probe amplification (MLPA) to determine the copy numbers of all exons. A heterozygous deletion that included exons 2-4 was identified in one of the patients. MLPA revealed that the other patient had two copies for all HMGCL exons. Paternal uniparental isodisomy of chromosome 1 was confirmed in this patient by microarray analysis. These findings indicate that MLPA is useful for the identification of genomic aberrations and mutations other than small-scale nucleotide alterations. To the best of our knowledge, this is the first study describing HMGCL deficiency caused by uniparental disomy.
authors: Medical Information Sciences Division, United Graduate School of Drug Discovery and Medical information Sciences, Gifu University, Gifu 501-1194, Japan.
authors: Yuka
authors: Y
authors: Aoyama
authors: Tokyo Women's Medical University Institute for Integrated Medical Sciences (TIIMS), Tokyo 162‑8666, Japan.
authors: Toshiyuki
authors: T
authors: Yamamoto
authors: Department of Pediatrics, Graduate School of Medicine, Gifu University, Gifu 501‑1194, Japan.
authors: Naomi
authors: N
authors: Sakaguchi
authors: Department of Pediatrics, Nihon University School of Medicine, Sapporo, Hokkaido 063‑0005, Japan.
authors: Mika
authors: M
authors: Ishige
authors: Department of Pediatrics and Clinical Research, NHO Hokkaido Medical Center, Sapporo, Hokkaido 063‑0005, Japan.
authors: Toju
authors: T
authors: Tanaka
authors: Department of Pediatrics, Takamatsu Red Cross Hospital, Takamatsu, Kagawa 760-0017, Japan.
authors: Tomoko
authors: T
authors: Ichihara
authors: Department of Pediatrics, Takamatsu Red Cross Hospital, Takamatsu, Kagawa 760-0017, Japan.
authors: Katsuaki
authors: K
authors: Ohara
authors: Department of Pediatrics, Takamatsu Red Cross Hospital, Takamatsu, Kagawa 760-0017, Japan.
authors: Hiroko
authors: H
authors: Kouzan
authors: Medical Information Sciences Division, United Graduate School of Drug Discovery and Medical information Sciences, Gifu University, Gifu 501-1194, Japan.
authors: Yasutomi
authors: Y
authors: Kinosada
authors: Department of Pediatrics, Graduate School of Medicine, Gifu University, Gifu 501‑1194, Japan.
authors: Toshiyuki
authors: T
authors: Fukao
mesh_terms: D020087:Alu Elements; D001483:Base Sequence; D002878:Chromosomes, Human, Pair 1 / Q000235:genetics* / Q000378:metabolism; D005091:Exons; D005260:Female; D006801:Humans; D007223:Infant; D008297:Male; D008661:Metabolism, Inborn Errors / Q000201:enzymology / Q000235:genetics* / Q000473:pathology; D060885:Multiplex Polymerase Chain Reaction; D007652:Oxo-Acid-Lyases / Q000172:deficiency* / Q000378:metabolism; D017384:Sequence Deletion; D024182:Uniparental Disomy / Q000235:genetics* / Q000473:pathology
chemical_list: D007652:Oxo-Acid-Lyases; C021410:3-hydroxy-3-methylglutaryl-coenzyme A lyase