Novel nanotechnology to concentrate and preserve Trypanosoma cruzi antigens in urine of Chagas disease patients co-infected with HIV virus

Y.E. Castro-Sesquen, R.H. Gilman, M. Reimer, C. Mejia, E. Valencia, L. Liotta, C. Bern and A. Luchini
George Mason University, US

Keywords: HIV/Chagas disease coinfection, nanotechnology, urine antigen detection

Summary:

Chagas disease, caused by Trypanosoma cruzi, is a severe opportunistic infection in immunosuppressed patients. The treatment of Chagas disease in these patients can be achieved most efficiently and it can be lifesaving if it is initiated soon after reactivation. T. cruzi antigens in urine of patients co-infected with T. cruzi/HIV were concentrated using hydrogel nanoparticles. 39 urine samples of HIV patients were evaluated: 20 co-infected with T. cruzi/HIV and 19 samples of patients without T. cruzi. N-isopropylacrylamide (NIPAm)-acrylic acid (AAc) nanoparticles were coupled with trypan blue. Urines were concentrated using the poly(NIPAm)/trypan blue nanoparticles. Antigens were detected by Western Blot. A detection limit of 0.8 ng/ml T. cruzi antigens was achieved. Two bands of 83 kDa and 18 kDa were detected in urine samples of patients with Chagas disease. The sensitivity of antigen detection was 90% (9/10, compared to PCR), and 80% (16/20, compared to ELISA). Antigen detection in urine were directly correlated to high levels of parasite DNA in blood, indicating that detection of T. cruzi antigens in urine can be an early predictor of reactivation of Chagas disease in HIV patients. Poly(NIPAm)/trypan blue nanoparticles protects H49 antigen from enzymatic degradation in model solutions and in urine samples.