Biomedical subjects
Publications and source records attributed to Duncan Graham.
Diels-Alder cycloaddition has been used to add a benzotriazole azo dye to a diene tagged oligonucleotide to generate unique SERRS signals at 10 attomoles.
Surface-enhanced resonance Raman scattering (SERRS) of a model derivative of TNT was detected using a microflow cell designed within the framework of the lab-on-a-chip concept, using only the analyte and readily available reagents. The SERRS substrate, silver colloid, was prepared in situ, on-chip, by borohydride reduction of silver nitrate. The silver colloid was imaged within the chip using a white light microscope in either transmission or, due to the high reflectivity of the colloid, reflection mode. A fine stream of colloid approximately 30 microm in width was formed in a 250-microm-wide channel at the point where the colloid preparation reagents met. The chip was designed to produce a concentrated stream of colloid within a laminar regime, such that particles did not readily disperse into the fluid. One result of this was to reduce the effective volume of analysis. Attempts to deliberately disrupt this stream with microstructured pillars, fabricated in the fluidic channels, were unsuccessful. The chip was also designed to have the appropriate dimensions for detection using a modern Raman microscope system, which collects scattering from a very small volume. A dye derived from TNT was used as a model analyte. Quantitative behavior was obtained over 4 orders of magnitude with a detection limit of 10 fmol. This performance is between 1 and 2 orders of magnitude better than that achieved using a macroflow SERRS cell. The technique has the added advantage that both reagent consumption and effluent production are greatly reduced, leading to reduced operating costs and a decreased environmental impact
Selective chemical functionalisation of 2,4,6-trinitrotoluene to a surface enhanced resonance Raman active species for sensitive detection.
The accurate detection of DNA sequences is essential for a variety of post human genome projects including detection of specific gene variants for medical diagnostics and pharmacogenomics. A specific DNA sequence detection assay based on surface-enhanced resonance Raman scattering (SERRS) and an amplification refractory mutation system (ARMS) is reported. Initially, generation of PCR products was achieved by using specifically designed allele-specific SERRS active primers. Detection by SERRS of the PCR products confirmed the presence of the sequence tested for by the allele-specific oligonucleotides. This lead directly to the multiplex genotyping of human DNA samples for the deltaF508 mutational status of the cystic fibrosis transmembrane conductance regulator gene using SERRS active primers in an ARMS assay. Removal of the unincorporated primers allowed fast and accurate analysis of the three genotypes possible in this system in a multiplex format without any separation of amplicons. The results indicate that SERRS can be used in modern genetic analysis and offers an opportunity for the development of novel assays. This is the first demonstration of the use of SERRS in multiplex genotyping and shows potential advantages over fluorescence as a detection technique with considerable promise for future development.
The synthesis of seven monoazo benzotriazole dyes for use in surface enhanced resonance Raman scattering, SERRS, is reported. The dyes are all capable of complexing to the silver surface used to provide the surface enhancement found in SERRS and hence act as 'model' analytes. One dye was examined in detail and showed a quantitative relationship between concentration and signal intensity.
Since the first use of DNA to identify the perpetrator of a murder in 1985, forensic science has witnessed dramatic changes in the field of human identification. The technology has altered by adopting novel methods developed originally for use in the field of medical genetics. Currently, millions of samples from blood, semen, hair and tissues are analyzed to determine the origin of the samples. The processes used at present rely on the separation of polymorphic DNA fragments by electrophoresis. Although rapid, this process represents a bottleneck in the automation of the process. Recent advances in chip-based techniques offer a rapid and highly automated solution, provided that the necessary DNA polymorphisms can be examined in this way. This review examines the immediate future of human identification and considers possible routes for future developments.