Detection of genetic variations in coagulopathy-related genes using ramified rolling circle amplification.

Abstract:

:We evaluated single nucleotide polymorphism (SNP) detection via a target-capture, C-probe ligation, and RAM assay in a single-blind comparison to clinical samples that had been tested with FDA-cleared tests for up to 4 different vascular disease-related SNPs. In the RAM assay circulizable linear probes (C- or padlock probes) were annealed directly to genomic DNA, processed on a largely automated platform, and ligated C-probes were amplified by real-time RAM. After allele determinations were made with the experimental system, the sample genotypes were unblinded and the experimentally determined genotypes were found to be completely consistent with the FDA-cleared test results. The methods and results presented here show that a combination of C-probes, automated sample processing, and isothermal RAM provides a robust, and specific, nucleic acid detection platform that is compatible with automated DNA sample preparation and the throughput requirements of the clinical laboratory.

journal_name

Biomed Res Int

authors

Smith JH,Cui M,Zhang DY,Beals TP,Ye F

doi

10.1155/2014/641090

subject

Has Abstract

pub_date

2014-01-01 00:00:00

pages

641090

eissn

2314-6133

issn

2314-6141

journal_volume

2014

pub_type

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