High resolution melting analysis for the detection of SLC25A13 gene mutations in Taiwan.
BACKGROUND
Citrin, encoded by SLC25A13 gene, is a mitochondrial solute transporter with a crucial role in urea, nucleotide and protein synthesis. SLC25A13 mutations cause two phenotypes, adult-onset type II citrullinemia and neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD). This study aimed to develop a high resolution melting (HRM) analysis for SLC25A13 mutation scanning and determine the carrier rate in Taiwan.


METHODS
DNAs from healthy subjects (n=479), and patients with hepatocellular carcinoma (HCC, n=100) and NICCD (n=5) were scanned in exons 6, 9, 11, 16, and 17 and parts of introns of SLC25A13 using HRM analysis. All mutations detected by HRM analysis were further confirmed by TaqMan method and/or direct sequencing.


RESULTS
In healthy subjects, seventeen carriers with mutants c.851_854del (n=10), c.1638_1660dup, c.615+5G>A (n=4), and two novel mutants, c.475C>T and c.1658G>A, were detected. The frequency of carriers was about 1/28. In patients with HCC, there were only 2 carriers with c.851_854del mutant. Patients with NICCD (n=5) diagnosed during 2007 and 2008, harbored compound heterozygous mutations c.851_854del/c.1177+1G>A, c.851_854del/c.1638_1660dup (n=2), c.851_854del/c.615+5G>A, and c.1638_1660dup/c.615+5G>A.


CONCLUSIONS
HRM analysis is a simple, rapid and robust method for detecting SLC25A13 mutations in clinical laboratories. SLC25A13 mutations may not be a major contributor to the pathogenesis of HCC in Taiwan.
Department of Medical Laboratory Science and Biotechnology, National Cheng Kung University, Tainan, Taiwan.
Jing-Ting
JT
Lin
Kwang-Jen
KJ
Hsiao
Chiung-Yu
CY
Chen
Cheng-Chung
CC
Wu
Shio-Jean
SJ
Lin
Yen-Yin
YY
Chou
Shu-Chu
SC
Shiesh
D006528:Carcinoma, Hepatocellular / Q000175:diagnosis / Q000235:genetics*; D002780:Cholestasis, Intrahepatic / Q000175:diagnosis / Q000235:genetics; D005838:Genotype; D006801:Humans; D008113:Liver Neoplasms / Q000175:diagnosis / Q000235:genetics*; D033681:Mitochondrial Membrane Transport Proteins / Q000235:genetics*; D009154:Mutation / Q000235:genetics*; D009691:Nucleic Acid Denaturation; D016133:Polymerase Chain Reaction; D013624:Taiwan; D044366:Transition Temperature
D033681:Mitochondrial Membrane Transport Proteins; C119629:SLC25A13 protein, human