title: 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.
abstract: 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.
authors: Department of Pediatrics, The First Affiliated Hospital, Jinan University, Guangzhou, Guangdong 510630, P.R. China.
authors: Han-Shi
authors: HS
authors: Zeng
authors: Department of Pediatrics, The First Affiliated Hospital, Jinan University, Guangzhou, Guangdong 510630, P.R. China.
authors: Wei-Xia
authors: WX
authors: Lin
authors: Department of Pediatrics, The First Affiliated Hospital, Jinan University, Guangzhou, Guangdong 510630, P.R. China.
authors: Shu-Tao
authors: ST
authors: Zhao
authors: Core Laboratory, The First Affiliated Hospital, Jinan University, Guangzhou, Guangdong 510630, P.R. China.
authors: Zhan-Hui
authors: ZH
authors: Zhang
authors: Biomedical Translational Research Institute, The First Affiliated Hospital, Jinan University, Guangzhou, Guangdong 510630, P.R. China.
authors: Heng-Wen
authors: HW
authors: Yang
authors: Department of Laboratory Science, The First Affiliated Hospital, Jinan University, Guangzhou, Guangdong 510630, P.R. China.
authors: Feng-Ping
authors: FP
authors: Chen
authors: Department of Pediatrics, The First Affiliated Hospital, Jinan University, Guangzhou, Guangdong 510630, P.R. China.
authors: Yuan-Zong
authors: YZ
authors: Song
authors: Biomedical Translational Research Institute, The First Affiliated Hospital, Jinan University, Guangzhou, Guangdong 510630, P.R. China.
authors: Zhi-Nan
authors: ZN
authors: Yin
mesh_terms: D001483:Base Sequence; D015415:Biomarkers; D020159:Citrullinemia / Q000175:diagnosis* / Q000235:genetics* / Q000628:therapy; D003001:Cloning, Molecular; D018076:DNA, Complementary; D005091:Exons; D023061:Gene Order; D005838:Genotype; D006801:Humans; D007223:Infant; D008214:Lymphocytes / Q000378:metabolism; D008297:Male; D033681:Mitochondrial Membrane Transport Proteins / Q000235:genetics*; D059645:Mutation Rate; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion
chemical_list: D015415:Biomarkers; D018076:DNA, Complementary; D033681:Mitochondrial Membrane Transport Proteins; C119629:SLC25A13 protein, human