Utilization of high-resolution melting analysis to screen patients with neonatal intrahepatic cholestasis caused by citrin deficiency.
- Author:
Peng-qiang WEN
1
;
Guo-bing WANG
;
Zhan-ling CHEN
;
Dong CUI
;
Xiao-hong LIU
;
Li-fang YING
;
Ping SONG
;
Quan YUAN
;
Shu-li CHEN
;
Jian-xiang LIAO
Author Information
- Publication Type:Journal Article
- MeSH: Anion Transport Proteins; genetics; Base Sequence; Calcium-Binding Proteins; deficiency; China; Citrullinemia; diagnosis; genetics; metabolism; DNA; chemistry; genetics; Genetic Predisposition to Disease; Genotype; Humans; Mitochondrial Proteins; genetics; Molecular Sequence Data; Mutation; Nucleic Acid Denaturation; Organic Anion Transporters; deficiency; Sensitivity and Specificity
- From: Chinese Journal of Medical Genetics 2012;29(2):167-171
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo assess the feasibility of high-resolution melting (HRM) analysis for screening patients with neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD).
METHODSBased on previous studies on SLC25A13 gene in Chinese patients with NICCD, four hotspot mutations (851del4, 1638ins23, IVS6+5G>A and IVS16ins3kb) were selected. Results of the HRM analysis was validated using 50 negative controls and 20 patients with NICCD whose genotypes were confirmed previously by direct sequencing. With the established protocol, 171 suspected patients were enrolled. Samples with abnormal melting curves were further validated by DNA sequencing.
RESULTSHRM analysis can accurately determine the genotypes of all negative controls and patients. The sensitivity and specificity of the technique reached 100% (70/70). The melting curves of samples with the same genotype were highly reproducible. In 171 suspected patients, seven NICCD patients were detected by HRM. Identified mutations have included one case of 851del4 homozygote, one case of IVS6+5G>A heterozygote, 3 cases of 851del4 heterozygotes, one case of [IVS6+5G>A]+[ 851del4] and one case of [1638ins23+IVS16ins3kb]+[1638ins23]. All mutations were subsequently confirmed by DNA sequencing.
CONCLUSIONHRM analysis is a convenient, high-throughput and rapid technique for the screening of NICCD patients.
