CHEMILUMINESCENT WESTERN BLOT SYSTEM FOR AIDS DIAGNOSIS

THE GOAL OF PHASE I IS TO ESTABLISH A CHEMILUMINESCENT DETECTION SYSTEM FOR AIDS WESTERN BLOTTING BASED ON THE CHEMILUMINESCENT SUBSTRATE FOR ALKALINE PHOSPHATASE, AMPPD. ONE ADVANTAGE OF THIS SYSTEM WILL BE INCREASED SENSITIVITY OVER EXISTING WESTERN BLOT TESTS, WHICH RESULT IN STAINED MEMBRANE STRIPS. ALSO, A PERMANENT RECORD OF RESULTS WILL BE PRODUCED DIRECTLY ON FILM. A MODEL WESTERN BLOT SYSTEM USING HUMAN SERUM TRANSFERRIN AND COMMERCIALLY AVAILABLE ANTIBODIES WILL BE USED TO DEVELOP A NEW LUMINESCENCE-AMPLIFYING SYSTEM THAT WILL BE NECESSARY FOR NITROCELLULOSE MEMBRANE. BLOCKING CONDITIONS AND SENSITIVITY WILL ALSO BE OPTIMIZED. THE LUMINESCENT AMPLIFYING SYSTEM WILL RESULT IN A CHEMILUMINESCENT EMISSIONWITH A MAXIMUM AT APPROXIMATELY 470 NM. THIS WAVELENGTH IS A GOOD MATCH FOR CAPTURING THE CHEMILUMINESCENT SIGNAL ON POLAROID INSTANT BLACK AND WHITE FILM OR X-RAY FILM. THE OPTIMAL CONDITIONS DEVELOPED WITH THE TRANSFERRIN MODEL SYSTEM WILL THEN BE APPLIED TO COMPONENTS OF A DUPONT HIV WESTERN BLOT KIT, USING THE KIT'S NONINFECTIOUS CONTROLS TO TEST THE PERFORMANCE AND SENSITIVITY OF THE CHEMILUMINESCENTSYSTEM. THE RESULTS OBTAINED ON INSTANT FILM WILL BE SUITABLE FOR ARCHIVAL STORAGE.