SYNTHETIC HYBRIDIZATION PROBES FOR HERPES SIMPLEX TYPE 2

THE LONG TERM GOAL OF THIS RESEARCH IS TO DEVELOP SYNTHETIC DNA HYBRIDIZATION PROBES TO DETECT VIRAL INFECTIONS. DNA HYBRIDIZATION HAS PROVEN HIGHLY EFFECTIVE IN DETECTING VIRUSES, BUT CURRENT METHODS FOR MAKING DNA HYBRIDIZATION PROBES ARE EXPENSIVE TO COMMERCIALIZE SINCE THEY DEPEND ON ENZYMATICALLY MODIFIED CLONED DNA. SYNTHETIC DNA PROBES, COMPLEMENTARY TO KNOWN SEQUENCES, ARE RELATIVELY INEXPENSIVE TO PRODUCE. THESE PROBES HYBRIDIZE RAPIDLY TO SPECIFIC TARGET SEQUENCES IN DNA OR RNA AND ARE MORE SENSITIVE TO MUTATIONS AND STRAIN DIFFERENCES THAN CLONED PROBES. OUR GOAL IS TO DEVELOP AN OLIGONUCLEOTIDE-BASED TEST TO DETECT HERPES SIMPLEX VIRUS TYPE 2 (HSV-2). PHASE I INVOLVES THE SYNTHESIS AND TESTING OF NORMAL AND CHEMICALLY MODIFIED OLIGONUCLEOTIDES COMPLEMENTARY TO THE HSV THYMIDINE KINASE GENE. PHASE I WILL DETERMINE THE SENSITIVITY AND SPECIFICITY OF OLIGOMER PROBES FOR FILTER BOUND HSV SEQUENCES COMPARED TO CLONED PROBES AND THEIR ABILITY TO DIFFERENTIATE HSV TYPE 1 AND 2. PHASE II WILL REFINE THE USE OF ISOTOPIC AND NON-ISOTOPIC PROBES TO DETECT HSV IN CLINICAL SAMPLES. ONCE SUCH PROBES ARE FULLY TESTED CLINICAL DIAGNOSTIC PROBES FOR OTHER SEQUENCES CAN BE DEVELOPED (E.G. HEPATITIS B VIRUS, EPSTEIN-BARR VIRUS, AND CYTOMEGALOVIRUS).