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Biomedical subjects

Brian D Warner

Publications and source records attributed to Brian D Warner.

2 recordsLinked to original sources

Online resistance monitoring during autometallographic enhancement of colloidal Au labels for DNA analysis.

DNA diagnostics at the point-of-care requires biosensors that rely on highly sensitive transducers and are producible at low cost. A promising candidate technology is based on direct electrical detection of autometallographically enhanced Au labeled analytes. We present a substantial improvement to the previously used method by introducing online DC resistance monitoring during the autometallographic enhancement process. Since multi-step enhancement, washing, drying, and measurement cycles are eliminated, our method takes the direct electrical detection method a step further to applicability in a point-of-care environment. The feasibility of the novel method is demonstrated by its application in a simple DNA hybridization assay and the analysis of a single nucleotide polymorphism (SNP) using allele-specific hybridization. Unequivocal discrimination of all possible base pairing combinations in the SNP assay has been achieved. The SNP assay in particular indicates the potential of the method for analyte quantification.

Biosensing Techniques↗

Single nucleotide polymorphism analysis by chip-based hybridization and direct current electrical detection of gold-labeled DNA.

Single nucleotide polymorphism (SNP) analysis at the point of care requires a low cost detection technology that is capable of miniaturization, multiplexing, and high sensitivity. Direct current electrical detection (DCED) of DNA following nanoparticle labeling and silver enhancement is a promising candidate technology for point-of-care diagnostics. In this work we present, for the first time, SNP analysis in PCR products from patient samples using DCED, taking this platform technology a step closer to practical application. We developed a silane functionalized polymer for coating of biochip surfaces. This polymeric coating is stable under harsh conditions and has exceptionally high binding capacity. Allele-specific oligonucleotide probes were immobilized on chips coated with this polymer. Biotinylated PCR products of the human cholesteryl ester transfer protein gene from different patients were hybridized to the chips, labeled with gold nanoparticles, and autometallographically enhanced. The chips were scanned for DC electrical resistance by applying movable electrodes to the surface. Eighteen of nineteen patient samples were assigned the correct genotype. Our results demonstrate that SNP analysis of patient samples is feasible with DCED.

Affinity Labels↗