上海交通大学学报（医学版）

›› 2011, Vol. 31 ›› Issue (7): 957-.doi: 10.3969/j.issn.1674-8115.2011.07.019

• 论著（临床研究） • 上一篇    下一篇

MLPA在常染色体显性遗传多囊肾病基因诊断中的应用

于国鹏1, 齐 隽1, 龙 飞2, 黄驿晨1, 钱小强1, 陶 炯2, 陈建华1   

  1. 1.上海交通大学 医学院附属新华医院泌尿外科, 上海 200092； 2.上海市儿科医学研究所细胞遗传室, 上海 200092
  • 出版日期:2011-07-28 发布日期:2011-07-27
  • 通讯作者: 陈建华, 电子信箱: hikaru_csa@hotmail.com。
  • 作者简介:于国鹏（1984—）, 男, 硕士生；电子信箱: yuguopeng22@gmail.com。
  • 基金资助:

    上海交通大学医学院附属新华医院院基金（09YJ02）

Application of MLPA in gene diagnosis of autosomal dominant polycystic kidney disease

YU Guo-peng1, QI Jun1, LONG Fei2, HUANG Yi-chen1, QIAN Xiao-qiang1, TAO Jiong2, CHEN Jian-hua1   

  1. 1.Department of Urology, Xinhua Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200092, China；2.Cytogenetics Laboratory, Shanghai Institute for Pediatric Research, Shanghai 200092, China
  • Online:2011-07-28 Published:2011-07-27
  • Supported by:

    Foundation from Xinhua Hospital, Shanghai Jiaotong University School of Medicine, 09YJ02

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摘要：

目的 探讨多重连接探针扩增技术（MLPA）在常染色体显性遗传多囊肾病（ADPKD）基因诊断中的应用。方法 采用MLPA对20例ADPKD患者的PKD1基因和PKD2基因进行检测。MLPA检测结果显示单一外显子扩增或疑似扩增者采用RT-PCR检测验证；MLPA检测结果显示单一外显子缺失或疑似缺失者采用PCR检测验证，存在扩增产物者进行测序验证。结果 MLPA检测显示：1例患者单一外显子缺失（PKD1 Exon40），5例单一外显子疑似缺失（PKD1 Exon1、PKD1 Exon25、PKD2 Exon8、PKD2 Exon8、PKD1 Exon25），3例单一外显子疑似扩增（PKD1 Exon6、PKD1 Exon7、PKD1 Exon7）。经RT-PCR检测验证，1例患者单一外显子扩增（PKD1 Exon6）；经PCR检测与测序验证，1例患者单一外显子错义突变（PKD1 Exon40），1例单一外显子缺失（PKD2 Exon8）。结论 MLPA为ADPKD的基因诊断提供了一种新的方法。

关键词: 常染色体显性遗传多囊肾病, 基因突变, 多重连接探针扩增技术

Abstract:

Objective To investigate the application of multiplex ligation-dependent probe amplification (MLPA) in the gene diagnosis of autosomal dominant polycystic kidney disease (ADPKD). Methods MLPA was employed to detect the PKD1 gene and PKD2 gene in 20 patients with ADPKD. Verification with RT-PCR was performed for those with single exon duplication or suspected duplication detected by MLPA. Those with single exon deletion or suspected deletion detected by MLPA were verified with PCR, and sequencing analysis was conducted in those with amplification products. Results One patient with single exon deletion (PKD1 Exon40), 5 patients with single exon suspected deletion (PKD1 Exon1, PKD1 Exon25, PKD2 Exon8, PKD2 Exon8 and PKD1 Exon25) and 3 patients with single exon suspected duplication (PKD1 Exon6, PKD1 Exon7 and PKD1 Exon7) were detected by MLPA. One patient with single exon duplication (PKD1 Exon6) was verified by RT-PCR, and one patient with single exon missense mutation (PKD1 Exon40) and one patient with single exon deletion (PKD2 Exon8) were verified by PCR and sequencing analysis. Conclusion MLPA may serve as a new method for gene diagnosis of ADPKD.

Key words: autosomal dominant polycystic kidney disease, gene mutation, multiplex ligation-dependent probe amplification