Integrated amplification microarray system in a lateral flow cell for warfarin genotyping from saliva.
Clin Chim Acta
; 429: 198-205, 2014 Feb 15.
Article
em En
| MEDLINE
| ID: mdl-24360850
ABSTRACT
BACKGROUND:
Genetic polymorphisms in the CYP2C9 and VKORC1 genes have been linked to sensitivity of the anticoagulant drug warfarin. The aim of this study is to demonstrate a method for warfarin sensitivity genotyping using gel element microarray technology in a simplified workflow from sample collection to analysis and detection.METHODS:
We developed an integrated amplification microarray system combining PCR amplification, target labeling, and microarray hybridization within a single, closed-amplicon "lateral flow cell" for genotyping three single nucleotide polymorphisms (SNPs) that influence warfarin response. We combined nucleic acid extraction of saliva using the TruTip technology together with the lateral flow cell assay and with fully automated array detection and analysis.RESULTS:
The analytical performance of the assay was tested using 20 genotyped human genomic DNA samples and found to be sensitive down to 330 input genomic copies (1 ng). A follow-up pre-clinical evaluation was performed with 65 blinded saliva samples and the genotyping results were in agreement with those determined by bidirectional sequencing.CONCLUSIONS:
Combined with the use of non-invasive saliva samples, rapid DNA extraction, the lateral flow cell, automatic imaging and data analysis provides a simple and fast sample-to-answer microarray test for warfarin sensitivity genotyping.Palavras-chave
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Base de dados:
MEDLINE
Assunto principal:
Saliva
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Varfarina
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Reação em Cadeia da Polimerase
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Análise de Sequência com Séries de Oligonucleotídeos
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Técnicas de Genotipagem
Idioma:
En
Ano de publicação:
2014
Tipo de documento:
Article