SLC25A13 cDNA cloning analysis using peripheral blood lymphocytes facilitates the identification of a large deletion mutation: Molecular diagnosis of an infant with neonatal intrahepatic cholestasis caused by citrin deficiency

• Published in: Molecular medicine reports Vol. 14; no. 6; pp. 5189 - 5194
• Main Authors: Zeng, Han-Shi, Lin, Wei-Xia, Zhao, Shu-Tao, Zhang, Zhan-Hui, Yang, Heng-Wen, Chen, Feng-Ping, Song, Yuan-Zong, Yin, Zhi-Nan
• Format: Journal Article
• Language: English
• Published: Greece D.A. Spandidos 01.12.2016; Spandidos Publications; Spandidos Publications UK Ltd
• Subjects: Age; Base Sequence; Biomarkers; Care and treatment; cDNA cloning; Children; Cholestasis; citrin deficiency; Citrullinemia - diagnosis; Citrullinemia - genetics; Citrullinemia - therapy; Clonal deletion; Cloning; Cloning, Molecular; Complementary DNA; Deoxyribonucleic acid; Development and progression; Diagnosis; Diseases; DNA; DNA polymerase; DNA, Complementary; Exons; Galactose; Gallbladder diseases; Gene deletion; Gene mutations; Gene Order; Genetic analysis; Genetic aspects; Genetic testing; Genotype; Health aspects; Hereditary diseases; Humans; Infant; Insertion; Laboratories; Lactose; Lymphocytes; Lymphocytes - metabolism; Male; Medical diagnosis; Mitochondrial Membrane Transport Proteins - genetics; Mutation; Mutation Rate; Neonates; Newborn babies; Pediatrics; Peripheral blood; peripheral blood lymphocytes; Polymerase chain reaction; Protein deficiency; Reverse transcription; Sequence Analysis, DNA; Sequence Deletion; SLC25A13; Beijing China; United States > US; China
• ISSN: 1791-2997; 1791-3004
• DOI: 10.3892/mmr.2016.5873

Neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD) is an autosomal recessive disorder resulting from biallelic mutations of the SLC25A13 gene. Due to the lack of well-recognized clinical or biochemical diagnostic criteria, the definitive diagnosis of this disease relies on the genetic analysis of SLC25A13 at present. As novel large deletion/insertion mutations of the SLC25A13 gene are difficult to detect using routine DNA analytic approaches, the timely diagnosis of patients with these types of mutations remains a challenge. The present study aimed to examine SLC25A13 mutations in an infant with a suspected diagnosis of NICCD. DNA was extracted from blood samples, and SLC25A13 mutations were examined by screening for high-frequency mutations and Sanger sequencing. Reverse transcription-polymerase chain reaction and cDNA cloning analyses were then performed using peripheral blood lymphocytes (PBLs) to identify the obscure mutation. The results demonstrated that the infant was heterozygous for a paternally-inherited mutation, c.851_854del4, and a maternally-inherited large deletion, c.1019_1177+893del, which has not been reported previously. A positive diagnosis of NICCD was made, and the infant responded favorably to a galactose-free and medium-chain triglyceride-enriched formula. The present study confirmed the effectiveness of this formula in NICCD therapy, enriched the SLC25A13 mutational spectrum and supported the feasibility of cDNA cloning analysis using PBLs as a molecular tool for facilitating the identification of large SLC25A13 deletions.