The use of an in-house modified double antibody sandwich ELISA to detect Aspergillus antigens in sera of immunosuppressed patients.
OBJECTIVE
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.


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.


CONCLUSIONS
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.
Samad
S A
SA
Department of Medical Microbiology and Immunology, Universiti Kebangsaan Malaysia Jalan Raja Muda Abdul Aziz, Kuala Lumpur, Malaysia.
Rahman
H A
HA
D000293:Adolescent; D000908:Antibodies, Fungal / Q000032:analysis*; D001228:Aspergillosis / Q000175:diagnosis* / Q000276:immunology; D001232:Aspergillus fumigatus / Q000276:immunology / Q000302:isolation & purification*; D002648:Child; D002675:Child, Preschool; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D016469:Fungemia / Q000175:diagnosis* / Q000276:immunology; D006801:Humans; D016867:Immunocompromised Host; D008297:Male; D008875:Middle Aged; D015203:Reproducibility of Results; D012189:Retrospective Studies; D012494:Sampling Studies; D012680:Sensitivity and Specificity
D000908:Antibodies, Fungal