The use of an in-house modified double antibody sandwich ELISA to detect Aspergillus antigens in sera of immunosuppressed patients.
AIM: The purpose of this study was to retrospectively detect Aspergillus antigens in sera obtained from immunocompromised host using an in-house modified double antibody sandwich ELISA. MATERIALS AND METHODS: The ELISA employed the use of polyclonal antibodies raised against a water-soluble (WS) mycelial component of a clinical isolate of Aspergillus fumigatus. Rabbit and guinea-pig anti-WS antibodies were used as the capture and detector antibodies respectively. This was followed by the addition of anti-guinea pig antibodies conjugated to horse radish peroxidase and orthophenylenediamine. RESULTS: Of 143 specimens obtained from 140 patients, only 5 (3.5%) specimens demonstrated the presence of antigen. The 5 antigen-positive sera were from three patients. One was a confirmed case of paranasal aspergillosis and the other two were probable cases of invasive aspergillosis. CONCLUSION: The clinical validity of the ELISA should be determined by prospective detection of Aspergillus antigens in multiple serum samples collected from proven cases of invasive aspergillosis.