A high-throughput population screening system for the estimation of genetic risk for type 1 diabetes:: An application for the TEDDY (The environmental determinants of diabetes in the young) study
A high-throughput population screening system for the estimation of genetic risk for type 1 diabetes:: An application for the TEDDY (The environmental determinants of diabetes in the young) study
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DOI：
10.1089/dia.2007.0229
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发表时间：
2007-10-01
影响因子：
5.4
通讯作者：
Ilonen, Jorma
中科院分区：
文献类型：
--
作者：
Kiviniemi, Minna;Hermann, Robert;Ilonen, Jorma
背景资料：在TEDDY（年轻人糖尿病的环境决定因素）研究中，患者资格是基于某些选定的1型糖尿病风险相关人类白细胞抗原DR-DQ基因型的存在。需要一种实用的筛查策略，在早期有效排除不合格的患者。此外，一个简单的，低成本的，快速的筛选系统是必不可少的主要步骤的风险评估，包括数以千计的samples.Methods：一个同质的基因分型系统，利用不对称聚合酶链反应（PCR）和随后的杂交等位基因特异性探针被设计为使用作为第一个筛选步骤。该检测方法相结合，进一步阐明了1型糖尿病的遗传风险，以筛选高风险individual.Results的方法：均质检测平台允许在一个工作日内对数百个样本进行分型。该测定的成本是最小的，并且与包括单独的PCR和杂交步骤的异质基因分型方法相比，动手时间的减少提供了相当大的改进。在第一步中使用的主要选择标准被证明是有效的，因为在随后的阶段分型的样品的数量显着reduced.Conclusions：所提出的检测系统提供了一个实用的方法，以低成本，大规模的筛选研究的一般出发点，数以千计的样品的快速筛选。
Background: In the TEDDY (The Environmental Determinants of Diabetes in the Young) study patient eligibility is based on the presence of some selected type 1 diabetes risk-associated human leukocyte antigen DR-DQ genotypes. A practical screening strategy was needed with efficient exclusion of ineligible patients at an early stage. Also, a simple, low-cost, and fast screening system was essential for the primary step of the risk assessment including thousands of samples.Methods: A homogeneous genotyping system utilizing an asymmetric polymerase chain reaction (PCR) and subsequent hybridization of allele-specific probes was designed to be used as the first screening step. This assay was combined with methods further elucidating the genetic risk of type 1 diabetes to screen for high-risk individuals.Results: The homogeneous assay platform allows the typing of hundreds of samples within one working day. The costs of the assay are minimal, and the reduction in hands-on time provides considerable improvements compared to the heterogeneous genotyping methods comprising separate PCR and hybridization steps. The primary selection criteria used in the first step proved to be efficient since the numbers of samples typed in subsequent stages were markedly reduced.Conclusions: The presented assay system provides a practical approach to the rapid screening of thousands of samples at low cost, a general starting point for large-scale screening studies.