Integrated amplification microarray system in a lateral flow cell for warfarin genotyping from saliva.

Autor: Sebastian T; Akonni Biosystems, Inc., 400 Sagner Avenue, Suite 300, Frederick, MD 21701, United States. Electronic address: tsebastian@akonni.com., Cooney CG; Akonni Biosystems, Inc., 400 Sagner Avenue, Suite 300, Frederick, MD 21701, United States., Parker J; Akonni Biosystems, Inc., 400 Sagner Avenue, Suite 300, Frederick, MD 21701, United States., Qu P; Akonni Biosystems, Inc., 400 Sagner Avenue, Suite 300, Frederick, MD 21701, United States., Perov A; Akonni Biosystems, Inc., 400 Sagner Avenue, Suite 300, Frederick, MD 21701, United States., Golova JB; Akonni Biosystems, Inc., 400 Sagner Avenue, Suite 300, Frederick, MD 21701, United States., Pozza L; DNA Genotek, Inc., 2 Beaverbrook Road, Kanata, Ontario K2K 1L1, Canada., Iwasiow RM; DNA Genotek, Inc., 2 Beaverbrook Road, Kanata, Ontario K2K 1L1, Canada., Holmberg R; Akonni Biosystems, Inc., 400 Sagner Avenue, Suite 300, Frederick, MD 21701, United States.
Jazyk: angličtina
Zdroj: Clinica chimica acta; international journal of clinical chemistry [Clin Chim Acta] 2014 Feb 15; Vol. 429, pp. 198-205. Date of Electronic Publication: 2013 Dec 17.
DOI: 10.1016/j.cca.2013.12.009
Abstrakt: 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.
(Copyright © 2013 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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