Research Project
3497541
Our goal is to develop a rapid, simple DNA hybridization system for use in clinical samples. Current methods for performing hybridizations (eg. nitrocellulose dot blots) are difficult to apply to crude samples and require protracted sample preparation. One promising solution is the hybridization "sandwich" assay in which a first probe bound to a solid support (i.e. affinity matrix) selects the target DNA from the sample. A second probe then identifies the bound target. Synthetic DNA affinity supports, based on a new Teflon-polyacrylamide DNA synthesis resin, can be used as the affinity matrix. Synthetic DNA probes may also be used as the second probe. Such oligonucleotide sandwich supports are relatively inexpensive to produce. Phase I involves the synthesis and testing of oligonucleotide affinity support matrices complementary to Herpes simplex virus (HSV). Phase I will determine the sensitivity and specificity of oligomer probes for HSV-DNA bound to the affinity support matrix and their ability to differentiate HSV subtypes in the presence of heterologous DNA. Phase II will refine the use of DNA affinity supports using isotopic and non-isotopic probes to detect viruses and bacteria in clinical samples. Once sandwich assays are fully tested, we will produce clinical diagnostic tests for viruses and bacteria.
