Antibodies detection employing solgel immobilized parasites

COPELLO GJ, DE MARZI MC, DESIMONE MF, MALCHIODI EL, DÍAZ LE

Mar del Plata, Buenos Aires, República Argentina.

Congreso; XLIII Reunión Anual Sociedad Argentina de Investigación en Bioquímica y Biología Molecular; 2007

Sociedad Argentina de Investigación en Bioquímica y Biología Molecular

Immunoflourescence (IFA) and Enzyme Immuno Assays (EIA) are useful diagnosis techniques for different diseases through specific antibodies (Abs) detection. Both techniques involve several fixation alternatives for immobilization of microorganisms, parasites or cellular fragments. The objective of this work was to generate a versatile system for antigen covalent attachment for the detection of serum Abs from mice infected with different pathogens. For this purpose the attachment of Trypanosoma cruzi and Leishmania guyanensisover a silicon oxide polymer covered surface was developed. The films were prepared using the sol-gel method. Standard microscope slides were coated with TEOS and APTES as polymeric precursors. These slides were used for indirect IFA and EIA analysis and both compared with the heat fixation technique. When acetone was used as the coating solvent IFA analysis, employing mouse infected sera, showed the fluorescence of attached parasites without matrix background interference. Similar results were observed when EIA techniques were evaluated. Thus, EIA and IFA are suitable methods for Abs detection from sera of infected mice when the antigen was covalent attached in soft conditions. Finally, the present immobilization method was able to maintain antigenic capability of attached cells allowing leading to homogeneous, ready to use and long lasting time coated slides.Trypanosoma cruzi and Leishmania guyanensisover a silicon oxide polymer covered surface was developed. The films were prepared using the sol-gel method. Standard microscope slides were coated with TEOS and APTES as polymeric precursors. These slides were used for indirect IFA and EIA analysis and both compared with the heat fixation technique. When acetone was used as the coating solvent IFA analysis, employing mouse infected sera, showed the fluorescence of attached parasites without matrix background interference. Similar results were observed when EIA techniques were evaluated. Thus, EIA and IFA are suitable methods for Abs detection from sera of infected mice when the antigen was covalent attached in soft conditions. Finally, the present immobilization method was able to maintain antigenic capability of attached cells allowing leading to homogeneous, ready to use and long lasting time coated slides.