Project Grant R44TR003549

POINT-OF-CARE RAPID RNA TEST FOR ZIKA - POINT-OF-CARE RAPID RNA TEST FOR ZIKA CROSSLIFE TECHNOLOGIES INC. PROJECT SUMMARY/ABSTRACT WE AIM TO DEVELOP A RAPID POINT-OF-TEST THAT DETECTS ZIKV AT THE POINT-OF-NEED IN < 30 MINUTES AND IS APPROPRIATE FOR NON-LABORATORY SETTINGS AND NON-TECHNICAL PERSONNEL. OUR PROPRIETARY TEST EXPLOITS A NOVEL PROBE REACTION CHEMISTRY THAT ALLOWS MULTIPLEXED DETECTION OF DNA OR RNA WITHOUT SAMPLE PURIFICATION, MAKING IT OPERABLE AS A SIMPLE, HAND-HELD TEST. PATIENT SAMPLES TO BE INPUT DIRECTLY INTO THE DEVICE WITHOUT PRE- PROCESSING AND A DISPOSABLE CARTRIDGE CARRY OUT AMPLIFICATION OF ALL TARGETS AND REPORTS VISIBLE RESULTS THAT CAN BE READ BY EYE. WE HAVE DEMONSTRATED THE FEASIBILITY OF A NOVEL AND SIMPLE DETECTION OF ZIKV USING TARA (TEMPLATED ASSISTED RAPID ASSAY). THE KEY GOALS OF THE PHASE 2 PROJECT ARE TO 1) DEVELOP AND EVALUATE AUTOMATED PAPER STRIP DEVICE AND TO 2) VALIDATE IT USING CLINICAL SAMPLES. WE WILL EVALUATE THE AUTOMATED PAPER STRIP DEVICE USING 200 SAMPLES FROM A NAVY-IRB APPROVED PROTOCOL THROUGH OUR CRADA COLLABORATION WITH U.S. NAVAL MEDICAL RESEARCH CENTER AND THE U.S. NAVAL MEDICAL RESEARCH UNIT SIX. WE EXPECT A SENSITIVITY (>90%) AND SPECIFICITY (>90%) TO THE QRT-PCR DATA ON THE SAME SAMPLES. THE SUCCESSFUL ACHIEVEMENT OF 1) TIME TO RESULT < 30 MIN, 2) SENSITIVITY > 90%, SPECIFICITY > 90% AND LOD < 500 COPIES/TEST, 3) > 90% USER FEEDBACK ON EASE OF USE NEED FOR CLIA WAIVER, 4) REQUIRES NO INSTRUMENTATION, POWER SOURCE, OR REFRIGERATION OF REAGENTS, AND 5) QUANTITATIVE CRITERIA FOR MEASURING THE OUTCOME OF EACH TASK ARE EXCEEDS OR EQUIVALENT TO THE RT-PCR MOLECULAR TEST.