Locked nucleic acid (LNA) probes in high-throughput genetic analysis:: Application to an assay for type 1 diabetes-related HLA-DQB1 alleles

被引：11

作者：

Kiviniemi, M

Nurmi, J

Lövgren, T

Ilonen, J

机构：

[1] Univ Turku, Dept Virol, FIN-20520 Turku, Finland

[2] Univ Turku, MediCity Res Lab, FIN-20520 Turku, Finland

[3] Univ Turku, Dept Biotechnol, FIN-20520 Turku, Finland

来源：

CLINICAL BIOCHEMISTRY | 2005年 / 38卷 / 11期

关键词：

homogeneous DNA assay; time-resolved fluorometry; locked nucleic acid; type; 1; diabetes;

D O I：

10.1016/j.clinbiochem.2005.08.001

中图分类号：

R446 [实验室诊断]; R-33 [实验医学、医学实验];

学科分类号：

1001 ;

摘要：

Objectives: In large-scale genetic screening, an assay that is reliable, fast and easy to perform, and straightforwardly adapted to new analytes is a necessity. We describe a one-step assay for analyzing HLA-DQB1 alleles which are associated with susceptibility to type 1 diabetes. Design and methods: The assay is based on asymmetric PCR amplification and a homogeneous hybridization method. The specificity of the probes was improved by substituting LNA (locked nucleic acid) for DNA at the critical bases. Results: The functionality of the LNA containing probes was found to be superior compared to probes consisting of DNA only. The homogeneous assay gave a correct genotyping result in 100% of the cases, which included both extracted DNA samples and blood samples dried on sample collection cards. Conclusion: This homogeneous approach provides a simple method to define disease risk associated with HLA alleles for large-scale screening projects. (C) 2005 The Canadian Society of Clinical Chemists. All rights reserved.

引用

页码：1015 / 1022

页数：8

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