Locked nucleic acid (LNA) probes in high-throughput genetic analysis: application to an assay for type 1 diabetes-related HLA-DQB1 alleles.
Clin Biochem
; 38(11): 1015-22, 2005 Nov.
Article
em En
| MEDLINE
| ID: mdl-16137668
ABSTRACT
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 ANDMETHODS:
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.
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Base de dados:
MEDLINE
Assunto principal:
Antígenos HLA-DQ
/
Testes Genéticos
/
Reação em Cadeia da Polimerase
/
Oligonucleotídeos Antissenso
/
Diabetes Mellitus Tipo 1
Idioma:
En
Ano de publicação:
2005
Tipo de documento:
Article