DMCA
ELISA for early diagnosis of histoplasmosis (2004)
BibTeX
@MISC{Zancopé-oliveira04elisafor,
author = {Rosely Maria Zancopé-oliveira and Allan Jefferson Guimarães and Claudia Vera Pizzini and Herbert Leonel and Matos Guedes and Priscila Costa Albuquerque and Jose ́ Mauro Peralta and Andrew John Hamilton and Rosely Maria},
title = {ELISA for early diagnosis of histoplasmosis},
year = {2004}
}
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Abstract
An ELISA was developed and evaluated as a method for detecting antibodies against glycosylated and deglycosylated histoplasmin (HMIN). Sera from patients with histoplasmosis, paracoccidioidomycosis, sporotrichosis, coccidioidomycosis, aspergillosis, cryptococcosis and healthy donors were tested by ELISA against purified, deglycosylated histoplasmin (ptHMIN) and compared with purified, native (i.e. glycosylated) histoplasmin (pHMIN). Although cross-reactivity was not abolished when ptHMIN was used in the test, it was reduced (pHMIN ELISA 93 % versus ptHMIN ELISA 96%). However, there were statistically significant differences between the sensitivities of these two methods for the detection of antibodies (pHMIN ELISA 57 % versus ptHMIN ELISA 92 %; P, 0.001) and between the efficiency of the methods (pHMIN ELISA 83% versus ptHMIN ELISA 95 %; P, 0.001). These parameters compare better than previously published data relating to the use of treated HMIN in diagnostic ELISAs. Some of the reactivities of serum samples were compared by immunoblotting using deglycosylated HMIN and by immunodiffusion using the crude antigen. The results demonstrated that cross-reactions with heterologous sera in both ELISAs could also be observed in immunoblotting and arose from shared protein epitopes. These data suggest that ELISA using deglycosylated HMIN is a very sensitive diagnostic method and, by using commercially available antigen, it can be easily standardized and performed faster thanpreviousWestern blot-based tests using the sameantigen. It provides auseful adjunct to existing methods of diagnosis that could be applied even in situations where laboratory facilities were relatively limited.
Keyphrases
versus pthmin elisa early diagnosis phmin elisa significant difference laboratory facility crude antigen serum sample heterologous serum sensitive diagnostic method healthy donor thanpreviouswestern blot-based test available antigen diagnostic elisa protein epitope auseful adjunct treated hmin
