Park Jason Y, Narayan Srinivas B, Bennett Michael J
Department of Pathology and Laboratory Medicine, University of Pennsylvania School of Medicine and the Hospital of the University of Pennsylvania, Philadelphia, 19104, USA.
Clin Chem Lab Med. 2006;44(9):1090-1. doi: 10.1515/CCLM.2006.196.
Carnitine palmitoyltransferase 1A (CPT1A) deficiency is a metabolic disorder that occurs at a key checkpoint of fatty acid metabolism. A new form of CPT1A deficiency caused by a mutation at nucleotide 1436 (C>T), resulting in an amino acid substitution of leucine for proline at position 479 (P479L), has been isolated in Canadian First Nations and Inuit populations. The present study offers a molecular method for assessing CPT1A 1436 (C>T) mutation status.
CPT1A-deficient fibroblasts from four patient fibroblast cell lines and ten patient peripheral blood spots were all analyzed by polymerase chain reaction (PCR) coupled to restriction endonuclease (RE) treatment. Genomic DNA was PCR-amplified and treated with an RE specific for normal DNA. CPT1A 1436 (C>T) mutations were identified by resistance to RE treatment.
The RE-PCR assay identified homozygosity for the 1436 (C>T) mutation in four fibroblast cell lines and nine blood spots with CPT1A enzyme deficiency. In addition, the assay identified one blood spot that corresponded to the heterozygous genotype.
RE-PCR assay for the 1436 (C>T) mutation provides a rapid assay for the diagnosis of CPT1A deficiency resulting from this mutation. The assay will have utility in screening populations with a high prevalence of this genotype.
肉碱棕榈酰转移酶1A（CPT1A）缺乏症是一种发生在脂肪酸代谢关键检查点的代谢紊乱疾病。在加拿大第一民族和因纽特人群中发现了一种由核苷酸1436（C>T）突变引起的新型CPT1A缺乏症，该突变导致第479位氨基酸由脯氨酸替换为亮氨酸（P479L）。本研究提供了一种评估CPT1A 1436（C>T）突变状态的分子方法。
对来自四个患者成纤维细胞系的CPT1A缺陷型成纤维细胞和十个患者外周血斑进行聚合酶链反应（PCR）结合限制性内切酶（RE）处理分析。对基因组DNA进行PCR扩增，并用针对正常DNA的RE进行处理。通过对RE处理的抗性鉴定CPT1A 1436（C>T）突变。
RE-PCR分析在四个成纤维细胞系和九个具有CPT1A酶缺乏的血斑中鉴定出1436（C>T）突变的纯合性。此外，该分析还鉴定出一个对应于杂合基因型的血斑。
针对1436（C>T）突变的RE-PCR分析为诊断由该突变导致的CPT1A缺乏症提供了一种快速检测方法。该检测方法将有助于筛查该基因型高流行人群。
