钠牛磺胆酸共转运多肽(NTCP)缺陷病是近几年新发现的一种遗传性胆汁酸代谢病，其确诊依赖SLC10A1基因分析。我们前期已在NTCP缺陷病患者中发现多种SLC10A1新变异类型，但其致病性有待实验证实。本项目应用重组子克隆、免疫荧光、Western blotting、Minigene剪接分析和双荧光素酶报告基因检测等技术，深入研究SLC10A1新变异的致病性；然后根据致病性新变异具体性质，建立相应的PCR、LA-PCR或PCR-RFLP分子诊断方法并应用于本病患者高危筛查；在以上研究基础上，进一步应用Sanger测序等方法在本病高危患儿中识别更多SLC10A1新变异类型，用于后续致病性研究。本项目可为阐明SLC10A1新变异致病性提供分子遗传学依据，为NTCP缺陷病确诊提供新分子标记和诊断方法，同时为深化SLC10A1变异谱特征的科学认识提供实验支撑，具有重要科学意义和良好应用前景。

Sodium taurocholate cotransporting polypeptide (NTCP) deficiency is a genetic disorder of bile acid metabolism newly-described in recent several years, and its definite diagnosis relies on SLC10A1 genetic analysis. In previous preliminary study, our team has identified a variety of novel SLC10A1 variants in patients with NTCP deficiency; however, the pathogenicity of these variants needs to be confirmed by in-depth laboratory investigations. This project will intensively explore the pathogenicity of the novel SLC10A1 variants by such methods as cloning of recombinant constructs, immunofluorescence, Western blotting, Minigene splicing analysis, and dual-luciferase report assay. And then, PCR, LA-PCR or PCR-RFLP molecular diagnostic approaches will be developed for corresponding novel SLC10A1 variants, and applied into the high-risk screening for patients with NTCP deficiency in clinical practices. Following that, more novel SLC10A1 variants will be identified for subsequent pathogenicity analysis, by a series of techniques such as Sanger sequencing in patients highly-suspected to have NTCP deficiency. This project will give molecular genetic evidences for the clarification of the pathogenicity of novel SLC10A1 variants, supply new molecular markers and diagnostic approaches for the definite diagnosis of NTCP deficiency, and provide experimental support for the in-depth understanding of the SLC10A1 variant spectrum, thus having important scientific value as well as promising application prospect.