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
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
复制标题
使用外周血淋巴细胞进行SLC25A13 cDNA克隆分析有助于识别大缺失突变：柠檬酸缺乏引起的新生儿肝内胆汁淤积婴儿的分子诊断
DOI：
10.3892/mmr.2016.5873
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发表时间：
2016-12-01
影响因子：
3.4
通讯作者：
Yin, Zhi-Nan
中科院分区：
文献类型：
--
作者：
Zeng, Han-Shi;Lin, Wei-Xia;Yin, Zhi-Nan
citrin缺乏导致的新生儿肝内胆汁淤积症（NICCD）是一种常染色体隐性遗传病，由SLC25A13基因的双等位基因突变引起。由于缺乏公认的临床或生化诊断标准，目前该疾病的确诊依赖于对SLC25A13的基因分析。由于SLC25A13基因的新型大片段缺失/插入突变难以用常规DNA分析方法检测，对这类突变患者的及时诊断仍然是一个挑战。本研究旨在对一名疑似NICCD的婴儿进行SLC25A13突变检测。从血液样本中提取DNA，通过高频突变筛查和桑格测序检测SLC25A13突变。然后利用外周血淋巴细胞（PBLs）进行逆转录 - 聚合酶链反应和cDNA克隆分析以确定不明突变。结果显示，该婴儿为父系遗传的c.851_854del4突变和母系遗传的此前未报道过的c.1019_1177 + 893del大片段缺失的杂合子。该婴儿被确诊为NICCD，且对无乳糖和富含中链甘油三酯的配方奶反应良好。本研究证实了该配方在NICCD治疗中的有效性，丰富了SLC25A13突变谱，并支持了将PBLs作为一种分子工具进行cDNA克隆分析以促进SLC25A13大片段缺失鉴定的可行性。
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_854de14, 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.