杂合子家族性高胆固醇血症患者低密度脂蛋白（LDL）受体基因第3和4外显子中的两个新突变。

Two novel mutations in exon 3 and 4 of low density lipoprotein (LDL) receptor gene in patients with heterozygous familial hypercholesterolemia.

作者信息

Khan Samia Perwaiz, Ghani Rubina, Ahmed Khwaja Zafar, Yaqoob Zia

机构信息

Department of Pharmacology, Ziauddin University, Karachi.

出版信息

J Coll Physicians Surg Pak. 2011 Jul;21(7):403-6.

DOI:

PMID:21777527

Abstract

OBJECTIVE

To determine the common mutation of low density lipoprotein receptor in hypercholesterolemia patients requiring screening for heterozygous familial hypercholesterolemia (HeFH) in Karachi.

STUDY DESIGN

Case-series.

PLACE AND DURATION OF STUDY

Dr. Ziauddin Hospital Laboratory and Dr. Rubina Ghani's Pathological and Molecular Laboratories, Karachi, for the PCR bench work from June 2008 to October 2009.

METHODOLOGY

All the patients selected for this study were from Dr. Ziauddin Hospital and National Institute of Cardiovascular Diseases. All the patients having high total cholesterol and LDL-cholesterol were included in this study with premature coronary artery diseases or a family history of hypercholesterolemia. Exclusion criteria included Diabetes mellitus, hypertension, renal disease, hypothyroidism and steroid therapy. After lipid profile with overnight fasting, DNA was extracted from whole blood collected in EDTA (ethylenediamine tetra acetic acid) tube and multiplex PCR (polymerase chain reaction) using forward and reverse primers of exons 3, 4, 9 and 14 of base pairs 162, 431, 550 and 496 respectively.

RESULTS

Out of total of 120 hypercholesterolemia cases, 42 patients were classical cases of HeFH (heterozygous familial hypercholesterolemia) with xanthomas, xanthelasmas and LDL-C > 160 mg/dl. The total cholesterol (260± 57 mg/dL) and LDL-C (192 ± 39 mg/dL ) of cases was significantly high as compared to, controls having total cholesterol (184 ± 27 mg/dL) and LDL-C (105 ± 22 mg/dL), p > 0.001. Two novel point mutations were noted in exon 3 and exon 4. The other 78 cases were probable with raised LDL-C (low density lipoprotein cholesterol) and family history of premature coronary heart diseases.

CONCLUSION

The frequency of HeFH was 35% classical and 65% probable cases out of total 120 hypercholesterolemia patients from two tertiary care hospitals in Karachi. The point mutation on exon 3 and exon 4 of LDLR gene was the most common. PCR is useful for the detection of large re-arrangements in the LDL-receptor gene and is a rapid and reliable method for diagnosis of familial hypercholesterolemia.

摘要

目的

确定卡拉奇需要筛查杂合子家族性高胆固醇血症（HeFH）的高胆固醇血症患者中低密度脂蛋白受体的常见突变。

研究设计

病例系列研究。

研究地点和时间

2008年6月至2009年10月，在卡拉奇的齐亚丁医院实验室以及鲁比娜·加尼医生的病理与分子实验室进行PCR实验操作。

方法

本研究选取的所有患者均来自齐亚丁医院和国家心血管疾病研究所。所有总胆固醇和低密度脂蛋白胆固醇水平高且患有早发性冠状动脉疾病或有高胆固醇血症家族史的患者均纳入本研究。排除标准包括糖尿病、高血压、肾脏疾病、甲状腺功能减退和类固醇治疗。空腹过夜进行血脂检测后，从采集于乙二胺四乙酸（EDTA）管中的全血中提取DNA，并使用分别对应第3、4、9和14外显子的162、431、550和496个碱基对的正向和反向引物进行多重聚合酶链反应（PCR）。

结果

在120例高胆固醇血症病例中，42例为HeFH（杂合子家族性高胆固醇血症）典型病例，伴有黄色瘤、睑黄瘤且低密度脂蛋白胆固醇（LDL-C）>160mg/dl。与总胆固醇为（184±27mg/dL）且低密度脂蛋白胆固醇为（105±22mg/dL）的对照组相比，病例组的总胆固醇（260±57mg/dL）和低密度脂蛋白胆固醇（192±39mg/dL）显著更高，p>0.001。在第3外显子和第4外显子中发现了两个新的点突变。其他78例可能存在低密度脂蛋白胆固醇（LDL-C）升高以及早发性冠心病家族史。