Research progress in the methodology and clinical application of low-density lipoprotein receptor gene mutation and function determination

doi:10.3969/j.issn.1673-8640.2021.12.019

Abstract

Abstract:

Low-density lipoprotein receptor（LDLR） is one type of cell surface glycoprotein,which plays a role in maintaining cholesterol metabolism balance in vivo. LDLR gene mutation can cause the decreasing,even the absence of LDLR on the cell surface,reduce the body's ability to metabolize cholesterol,lead to elevated plasma low-density lipoprotein cholesterol（LDL-C） level and deposition in different tissues and organs,and cause a variety of lipid metabolism disorders. Accurate and rapid determination of LDLR gene mutations and analysis of changes in LDLR function are essential for the diagnosis and differential diagnosis of familial hypercholesterolemia（FH）. This review focuses on the progress in the methodology and clinical application of LDLR gene mutation and function determination,in order to raise awareness and concern of its importance in early diagnosis and intervention of FH patients.

Key words: Low-density lipoprotein receptor, Low-density lipoprotein cholesterol, Lipid metabolism, Gene mutation

CLC Number:

R446.1

ZHU Yonglin, WANG Lüya, YAN Shengkai. Research progress in the methodology and clinical application of low-density lipoprotein receptor gene mutation and function determination[J]. Laboratory Medicine, 2021, 36(12): 1287-1291.

Figures/Tables 3

References 30

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检测方法	原理	优点	缺点	应用示例
Sanger测序	利用双脱氧核苷酸可终止聚合酶联反应的原理	可检测长序列和5%的小等位基因片段;基因测序金标准;样本少时效益高	样本多时效益低;成本高;速度慢	225例受试者中发现了LDLR基因的7个致病突变和5个良性突变^[10]。
NGS	通过捕捉新添加碱基携带的特殊标记来确定DNA序列	高通量;可同时测序多个基因;可发现新变异;测序深度、灵敏度高	样本少时效益低;需进行生物信息学分析	148例阳性突变患者中发现了77例LDLR基因杂合型突变和5例LDLR基因纯合型突变^[12]。
MLPA	在靶序列变异位点两侧设计1对探针,经杂交、连接、聚合酶联反应扩增、产物分离及数据分析后得出结果	CNV的金标准;简单、成本低;突变检测阴性后使用	仅针对LDLR基因;不能检测单细胞和未知位点;对探针附近新的良性多态性敏感	在新加坡人群中检出4例大的LDLR基因新重排突变^[13]。

检测方法	原理	优点	缺点	应用示例
Sanger测序	利用双脱氧核苷酸可终止聚合酶联反应的原理	可检测长序列和5%的小等位基因片段;基因测序金标准;样本少时效益高	样本多时效益低;成本高;速度慢	225例受试者中发现了LDLR基因的7个致病突变和5个良性突变^[10]。
NGS	通过捕捉新添加碱基携带的特殊标记来确定DNA序列	高通量;可同时测序多个基因;可发现新变异;测序深度、灵敏度高	样本少时效益低;需进行生物信息学分析	148例阳性突变患者中发现了77例LDLR基因杂合型突变和5例LDLR基因纯合型突变^[12]。
MLPA	在靶序列变异位点两侧设计1对探针,经杂交、连接、聚合酶联反应扩增、产物分离及数据分析后得出结果	CNV的金标准;简单、成本低;突变检测阴性后使用	仅针对LDLR基因;不能检测单细胞和未知位点;对探针附近新的良性多态性敏感	在新加坡人群中检出4例大的LDLR基因新重排突变^[13]。

类别	检测目的	检测方法	检测原理	应用情况
荧光分析法	LDLR的表达	抗体标记法	FACS检测LDLR抗体的荧光强度	常用
		间接免疫荧光法	FACS检测荧光二抗的荧光强度	常用
		免疫印迹法	凝胶图像处理系统分析条带强度	常用
	LDLR结合LDL的能力	FACS检测法	FACS定量检测LDLR结合LDL的量	常用
		CLSM检测法	CLSM可视化观察LDLR结合LDL	常用
放射性分析法	LDLR的表达	放射性物质标记法	化学发光法检测细胞内¹²⁵I-IgG-C7的放射性	常用（但正在被荧光分析法取代）
放射性分析法	LDLR结合LDL的能力	放射性物质标记法	化学发光法检测肝素的放射性	常用（但正在被荧光分析法取代）

类别	检测目的	检测方法	检测原理	应用情况
荧光分析法	LDLR的表达	抗体标记法	FACS检测LDLR抗体的荧光强度	常用
		间接免疫荧光法	FACS检测荧光二抗的荧光强度	常用
		免疫印迹法	凝胶图像处理系统分析条带强度	常用
	LDLR结合LDL的能力	FACS检测法	FACS定量检测LDLR结合LDL的量	常用
		CLSM检测法	CLSM可视化观察LDLR结合LDL	常用
放射性分析法	LDLR的表达	放射性物质标记法	化学发光法检测细胞内¹²⁵I-IgG-C7的放射性	常用（但正在被荧光分析法取代）
放射性分析法	LDLR结合LDL的能力	放射性物质标记法	化学发光法检测肝素的放射性	常用（但正在被荧光分析法取代）