DIAGNOSTICS SPECIFIC FOR CHAGAS DISEASE, PHASE I

THE RESEARCH IN THIS APPLICATION WILL BE USED IN THE DEVELOPMENT OF NEW DIAGNOSTIC REAGENTS FOR CHAGAS' DISEASE. IT IS ESTIMATED THAT 10-12 MILLION PEOPLE HAVE THE DISEASE; 200 MILLION PEOPLE ARE EXPOSED TO THE REDUVIID BUG THAT IS THE VECTOR FOR THE CAUSITIVE AGENT, "TRYPONASOMA CRUZI". CURRENT SEROLOGICAL DIAGNOSTICS ARE INSENSITIVE AND CROSS REACT WITH OTHER PARASITIC DISEASES (SUCH AS LEISHMANISIS). THE FOCUS OF PHASE I WILL BE IDENTIFICATION OF PROTEIN ANTIGENS (AGS) THAT REACT SPECIFICALLY WITH SERUM FROM CHAGASTIC INDIVIDUALS. "T. CRUZI" ORGANISMS WILL BE RADIOACTIVELY LABELED AND USED IN IMMUNOPRECIPITATIONS AND WESTERN BLOTS. PRELIMINARY DATA SUGGEST A 90 KDA PROTEIN REACTS WITH CHAGASIC SERUM BUT NOT WITH SERA FROM INDIVIDUALS THAT HAVE LEISHMANIA. MRNA WILL BE ISOLATED THAT CAN BE TRANSLATED "IN VITRO" INTO THIS PROTEIN (VERIFIED BY V8 DIGESTS AND IMMUNOPRECIPITATIONS). CDNA WILL BE MADE FROM PURIFIED POLY A+ MRNA AND THE DIAGNOSTIC ANTIGEN CDNA CLONE WILL BE IDENTIFIED. IN PHASE II OF THE PROJECT, THE DIAGNOSTIC ANTIGEN CDNA CLONE WILL BE SUBCLONED, AND EXPRESSED IN BACTERIA OR YEAST. THIS WILL PROVIDE AN INEXPENSIVE SUPPLY OF ANTIGEN FOR USE IN SPECIFIC DIAGNOSIS OF CHAGAS' DISEASE. THIS RESEARCH PLAN IS BEING SUBMITTED TO THE MICROBIOLOGY AND INFECTIOUS DISEASE PROGRAM AT THE NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASE AS DESCRIBED IN PHS OMNIBUS SOLICITATION FOR SBIR PROGRAM.