Solid phase immobilized minor groove binder probes

DESCRIPTION (provided by applicant): Bacterial and viral meningitis and encephalitis is a major cause of morbidity and mortality in all societies of the world. The rapid progression of symptoms and potentially devastating effect of these diseases necessitate early recognition and immediate treatment. The large number of potential organisms involved, the fact that some infections are self-limiting while others may have potentially fatal outcome, complicates diagnosis and treatment. The goal of the proposed work is to develop an assay that will simultaneously detect all the common meningitis and encephalitis pathogens typically seen in a medical facility in the US. The potential pathogens will be amplified in a multiplexed PCR from a single cerebral spinal fluid (CSF) sample and detected on an array containing pathogen-specific probes that fluoresce on hybridization. In Phase I we have shown that a Pleiades probe immobilized to a hydrogel electronic NanoChip array cartridge fluoresces when hybridized to its complementary amplified target with signal to background ratios significantly better than any comparable immobilized probe. It was shown that the immobilized probe could distinguish between match, single-mismatch and double-mismatched amplified targets. In Phase II the common meningitis and encephalitis pathogens responsible for disease in the US will be identified. Primer pairs will be developed for amplification of each meningitis and encephalitis target, multiplexed to allow the detection of each target in a single CSF amplified sample. The proposed Phase II work will allow the simultaneous detection of about 90% of the pathogens typically seen in a US medical facility in contrast to about 45% of the organisms currently confirmed clinically. The proposed meningitis and encephalitis panel will be the first available in a clinical laboratory to assist diagnosis and treatment. The goal of the proposed Phase II project is to develop an array for the rapid detection of the common pathogens involved in meningitis and encephalitis in the US. This will enable the rapid differentiate of organisms that cause self-limiting disease from those with potentially fatal outcome.