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使用磁性/发光纳米颗粒对多囊肾病(PKD)基因突变进行快速定量DNA分析。

Rapid and quantitative DNA analysis of genetic mutations for polycystic kidney disease (PKD) using magnetic/luminescent nanoparticles.

作者信息

Son Ahjeong, Dhirapong Amy, Dosev Dosi K, Kennedy Ian M, Weiss Robert H, Hristova Krassimira R

机构信息

Department of Land, Air, and Water Resources, University of California, Davis, 1 Shields Avenue, Davis, CA 95616, USA.

出版信息

Anal Bioanal Chem. 2008 Apr;390(7):1829-35. doi: 10.1007/s00216-008-1892-4. Epub 2008 Feb 7.

Abstract

Rapid and accurate detection of genetic mutations based on nanotechnology would provide substantial advances in detection of polycystic kidney disease (PKD), a disease whose current methods of detection are cumbersome due to the large size and duplication of the mutated gene. In this study, a nanotechnology-based DNA assay was developed for detection of SNPs (single nucleotide polymorphisms) in a feline autosomal dominant PKD (ADPKD) model which can readily be adapted to diagnosis of human ADPKD type 1. Europium and terbium phosphors were doped into gadolinium crystal hosts with a magnetic core, providing stable luminescence and the possibility of magnetic manipulations in a solution-based assay. A hybridization-in-solution DNA assay was optimized for feline PKD gene SNP detection using genomic DNA extracted from feline kidney tissue and blood. This assay showed a substantial differentiation between PKD and control specimens. The nanotechnology-based DNA assay is attractive from the viewpoint of rapid availability, simple methodology, and cost reduction for clinical use to detect mutations involved in human ADPKD and other genetic diseases.

摘要

基于纳米技术快速准确地检测基因突变,将在多囊肾病(PKD)的检测方面取得重大进展。多囊肾病目前的检测方法因突变基因体积大且存在重复而繁琐。在本研究中,开发了一种基于纳米技术的DNA检测方法,用于检测猫常染色体显性多囊肾病(ADPKD)模型中的单核苷酸多态性(SNP),该模型可轻松应用于人类1型ADPKD的诊断。将铕和铽荧光粉掺杂到具有磁芯的钆晶体基质中,在基于溶液的检测中提供稳定的发光以及进行磁操作的可能性。使用从猫肾组织和血液中提取的基因组DNA,对用于猫PKD基因SNP检测的溶液杂交DNA检测方法进行了优化。该检测方法在PKD样本和对照样本之间显示出显著差异。从快速可用性、方法简单以及降低临床检测人类ADPKD和其他遗传疾病相关突变的成本等角度来看,基于纳米技术的DNA检测方法颇具吸引力。

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