ENZYME TAGGING OF MALIGNANT CERVICAL EPITHELIAL CELLS

THE LONG-TERM AIM OF THIS PROJECT IS THE DEVELOPMENT OF AN AUTOMATED SCREENING INSTRUMENT (REAGENTS INTEGRATED WITH INSTRUMENTATION) FOR PAP SMEARS. PHASE I WILL BE USED TO DEMONSTRATE THE FEASIBILITY OF USING 2 MONOCLONAL ANTIBODIES AGAINST 2 ENZYMES AS THE KEY REAGENTS IN THE SYSTEM. BRIEFLY THE PAP SMEAR WILL BE ALLOWED TO INTERACT WITH TWO MONOCLONAL ANTIBODIES EACH CARRYING A DISTINCTIVE FLOURESCENT REPORTER MOLECULAR. THESE ANTIBODIES HAVE BEEN SELECTED TO IDENTIFY AND QUANTITATE LACTATED DEHYDROGENASE (LD) AND ADENYL CYCLASE (AC) RESPECTIVELY. LD OCCURS IN HIGHER THAN NORMAL, AC IN LOW TO NORMAL CONCENTRATIONS IN NEOPLASTIC CELLS. A FLUOROMETER CAPABLE OF READING TWO DISCREET WAVELENGTHS SIMULTANEOUSLY, CAN THEREFORE DEVELOP A NUMERICAL RATIO, WHICH WILL ASSESS THE DEGREE TO WHICH NEOPLASIA IS REPRESENTED IN A GIVEN SLIDE. BASED ON THESE RATIOS, NORMAL SLIDES (85-90% OF ALL THE SLIDES) WILL NEED NO FURTHER EVALUATION, ABNORMAL SLIDES ONLY, WILL NEED TO BE READ BY THE CYTOTECH/CYTOLOGIST.