Project Grant R01EY036137

RAPID DIAGNOSIS OF OCULAR INFECTIONS USING A SENSITIVE AND HIGHLY MULTIPLEXED BROAD PATHOGEN DETECTION ASSAY - PROJECT SUMMARY TIMELY AND ACCURATE DIAGNOSIS OF OCULAR INFECTIONS IS CRITICAL TO SUPPORT EARLY EFFECTIVE THERAPY AND SAVE VISION. SERIOUS INFECTIONS SUCH AS INFECTIOUS KERATITIS, UVEITIS AND ENDOPHTHALMITIS ARE LEADING CAUSES OF BLINDNESS GLOBALLY. THESE INFECTIONS ARE CAUSED BY A WIDE RANGE OF PATHOGENS, WHICH CAN OFTEN PRESENT WITH OVERLAPPING FEATURES AND ARE DIFFICULT TO DIFFERENTIATE CLINICALLY. LABORATORY-BASED DIAGNOSIS IS AN IMPORTANT COMPONENT OF CLINICAL MANAGEMENT TO GUIDE EFFICIENT DEPLOYMENT OF THERAPIES. HOWEVER, BECAUSE CURRENT METHODS EMPLOYED FOR PATHOGEN DETECTION ARE OUTDATED, TIME-CONSUMING AND LACK SENSITIVITY, PATIENTS CAN BE TREATED FOR PROLONGED PERIODS WITH COMBINATIONS OF EMPIRICAL BROAD-SPECTRUM THERAPIES THAT ARE NOT TAILORED FOR AN INDIVIDUAL, MAY BE INEFFECTIVE, CAUSE NEEDLESS OCULAR TOXICITY, AND ARE BECOMING COMPROMISED BY THE EMERGENCE OF ANTIMICROBIAL RESISTANCE. RAPID PRECISION DIAGNOSTIC TESTS ARE CRITICALLY NEEDED TO HELP GUIDE THERAPY AND IMPROVE PATIENT OUTCOMES. TO ADDRESS THIS CRITICAL AND URGENT NEED, WE ARE DEVELOPING NOVEL AND HIGHLY MULTIPLEXED TARGETED ASSAYS FOR COMPREHENSIVE DETECTION OF PATHOGENS FROM OCULAR SPECIMENS WITH HIGH ACCURACY AND SPEED. THIS INNOVATIVE APPROACH LEVERAGES THE ADVANTAGES OF THE CUTTING-EDGE NANOSTRING TECHNOLOGY, WHICH ENABLES RAPID AND SIMULTANEOUS DETECTION AND QUANTIFICATION OF HUNDREDS OF DIFFERENT DNA OR RNA MOLECULES WHILE MAINTAINING EXCELLENT SENSITIVITY PER TARGET. IN PRELIMINARY STUDIES, WE DEMONSTRATED THAT OUR NOVEL DIAGNOSTIC APPROACH CAN DETECT THE MOST IMPORTANT ETIOLOGIES OF SIGHT-THREATENING INFECTIONS IN A HIGHLY MULTIPLEXED FORMAT, WITH HIGH SENSITIVITY AND SPECIFICITY, AND IN A TIMELY MANNER, REDUCING THE TIME FROM SAMPLE TO ANSWER FROM DAYS TO 12 HOURS. WITH PROOF-OF-PRINCIPLE ESTABLISHED AND ANALYTICAL VALIDATIONS THOROUGHLY PERFORMED FOR AN INITIAL PANEL ABLE TO SIMULTANEOUSLY DETECT 48 TARGETS IN A SINGLE REACTION, WE ARE NOW AIMING TO 1) FURTHER DEVELOP, EXPAND, AND VALIDATE OUR PANEL WITH THE GOAL OF COVERING ADDITIONAL PATHOGENS OF RELEVANCE IN KERATITIS WITH BROADER GEOGRAPHICAL COVERAGE AND APPLICABILITY, AND 2) LAY THE GROUNDWORK NECESSARY TO TRANSLATE THIS TECHNOLOGY INTO PRACTICE AND FACILITATE ITS CLINICAL IMPLEMENTATION BY PERFORMING EXTENSIVE CLINICAL VALIDATIONS USING OUR EXISTING LARGE BIOREPOSITORIES OF SPECIMENS COLLECTED FROM PATIENTS PRESENTING WITH INFECTIOUS KERATITIS, UVEITIS AND ENDOPHTHALMITIS, AND ADDITIONAL SAMPLES THAT WILL CONTINUE TO BE COLLECTED IN THE FIRST YEARS OF THE PROJECT. THIS PROPOSAL WILL RESULT IN A TECHNICALLY NOVEL AND CLINICALLY USEFUL DIAGNOSTIC APPROACH THAT WILL ADVANCE THE DIAGNOSIS OF INFECTIONS ASSOCIATED WITH GREAT OCULAR MORBIDITY GLOBALLY, USING AN ASSAY THAT CAN DETECT ~95% OF THE ETIOLOGIES OF SIGHT-THREATENING INFECTIONS IN ONLY 12 HOURS, AND WITH HIGH ACCURACY, INCLUDING DIFFICULT TO CULTURE AND POTENTIALLY UNCULTURABLE PATHOGENS, MAKING A SINGLE-REACTION DIAGNOSTIC TEST COMPREHENSIVE AT AN UNPRECEDENTED LEVEL. THIS IS EXPECTED TO TRANSFORM OCULAR INFECTION TREATMENT BY SUPPORTING PROMPT INITIATION OF THERAPY WITH AN EFFECTIVE DRUG AT EARLY STAGES OF THE INFECTION, HELPING TO SAVE VISION AND PROMOTING ANTIMICROBIAL DE-ESCALATION TO REDUCE THE OVERUSE OF EMPIRICAL THERAPIES AND LIMIT THE SPREAD OF ANTIMICROBIAL RESISTANCE.