Identification of a Large SLC25A13 Deletion via Sophisticated Molecular Analyses Using Peripheral Blood Lymphocytes in an Infant with Neonatal Intrahepatic Cholestasis Caused by Citrin Deficiency (NICCD): A Clinical and Molecular Study.
Identification of a Large SLC25A13 Deletion via Sophisticated Molecular Analyses Using Peripheral Blood Lymphocytes in an Infant with Neonatal Intrahepatic Cholestasis Caused by Citrin Deficiency (NICCD): A Clinical and Molecular Study.
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DOI：
10.1155/2016/4124263
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发表时间：
2016
影响因子：
--
通讯作者：
Song YZ
中科院分区：
文献类型：
--
作者：
Zheng QQ;Zhang ZH;Zeng HS;Lin WX;Yang HW;Yin ZN;Song YZ
背景Citrin缺乏引起的新生儿肝内胆汁淤积症（NICCD）是一种由SLC 25 A13双等位基因突变引起的孟德尔遗传病，SLC 25 A13基因分析是确诊的必要条件。然而，传统的SLC 25 A13分析不能检测所有的突变，尤其是不明显的大插入/缺失。目的探讨1例NICCD患儿SLC 25 A13基因突变。方法.提取基因组DNA，筛选SLC 25 A13基因的4个高频突变，然后对所有18个外显子及其侧翼序列进行桑格测序。随后，进行cDNA克隆、SNP分析和半定量PCR以鉴定隐蔽突变。结果筛选出一个母系遗传的突变IVS 16 ins 3 kb，然后通过cDNA克隆，发现了父系遗传的具有外显子5跳跃的选择性剪接变体（ASVs）。通过对SLC 25 A13基因第5外显子附近的基因组DNA的深入研究，发现了一个c.329-1687_c.468+ 3865 del 5692 bp的大片段缺失，该缺失在其他文献中从未报道过。结论. 1例NICCD患者确诊为IVS 16 ins 3 kb和c.329-1687_c.468+ 3865 del 5692 bp复合杂合子。大的缺失丰富了SLC 25 A13突变谱，其鉴定支持了cDNA克隆分析，沿着其他分子工具如半定量PCR，可以提供有价值的线索，有助于确定模糊的SLC 25 A13缺失的概念。
Background. Neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD) is a Mendelian disorder arising from biallelic SLC25A13 mutations, and SLC25A13 genetic analysis was indispensable for its definite diagnosis. However, conventional SLC25A13 analysis could not detect all mutations, especially obscure large insertions/deletions. This paper aimed to explore the obscure SLC25A13 mutation in an NICCD infant. Methods. Genomic DNA was extracted to screen for 4 high-frequency SLC25A13 mutations, and then all 18 exons and their flanking sequences were analyzed by Sanger sequencing. Subsequently, cDNA cloning, SNP analyses, and semiquantitative PCR were performed to identify the obscure mutation. Results. A maternally inherited mutation IVS16ins3kb was screened out, and then cDNA cloning unveiled paternally inherited alternative splicing variants (ASVs) featuring exon 5 skipping. Ultimately, a large deletion c.329-1687_c.468+3865del5692bp, which has never been described in any other references, was identified via intensive study on the genomic DNA around exon 5 of SLC25A13 gene. Conclusions. An NICCD patient was definitely diagnosed as a compound heterozygote of IVS16ins3kb and c.329-1687_c.468+3865del5692bp. The large deletion enriched the SLC25A13 mutation spectrum, and its identification supported the concept that cDNA cloning analysis, along with other molecular tools such as semiquantitative PCR, could provide valuable clues, facilitating the identification of obscure SLC25A13 deletions.