ELISA for early diagnosis of histoplasmosis.
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 than previous Western blot-based tests using the same antigen. It provides a useful adjunct to existing methods of diagnosis that could be applied even in situations where laboratory facilities were relatively limited.
Guimarães
Allan Jefferson
AJ
Instituto de Pesquisa Clínica Evandro Chagas, Fundação Oswaldo Cruz, Rio de Janeiro, Brazil 2Instituto de Microbiologia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil 3St John's Institute of Dermatology, Guy's Hospital, King's College, London, UK.
Pizzini
Claudia Vera
CV
Instituto de Pesquisa Clínica Evandro Chagas, Fundação Oswaldo Cruz, Rio de Janeiro, Brazil 2Instituto de Microbiologia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil 3St John's Institute of Dermatology, Guy's Hospital, King's College, London, UK.
de Matos Guedes
Herbert Leonel
HL
Instituto de Pesquisa Clínica Evandro Chagas, Fundação Oswaldo Cruz, Rio de Janeiro, Brazil 2Instituto de Microbiologia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil 3St John's Institute of Dermatology, Guy's Hospital, King's College, London, UK.
Albuquerque
Priscila Costa
PC
Instituto de Pesquisa Clínica Evandro Chagas, Fundação Oswaldo Cruz, Rio de Janeiro, Brazil 2Instituto de Microbiologia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil 3St John's Institute of Dermatology, Guy's Hospital, King's College, London, UK.
Peralta
José Mauro
JM
Instituto de Pesquisa Clínica Evandro Chagas, Fundação Oswaldo Cruz, Rio de Janeiro, Brazil 2Instituto de Microbiologia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil 3St John's Institute of Dermatology, Guy's Hospital, King's College, London, UK.
Hamilton
Andrew John
AJ
Instituto de Pesquisa Clínica Evandro Chagas, Fundação Oswaldo Cruz, Rio de Janeiro, Brazil 2Instituto de Microbiologia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil 3St John's Institute of Dermatology, Guy's Hospital, King's College, London, UK.
Zancopé-Oliveira
Rosely Maria
RM
Instituto de Pesquisa Clínica Evandro Chagas, Fundação Oswaldo Cruz, Rio de Janeiro, Brazil 2Instituto de Microbiologia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil 3St John's Institute of Dermatology, Guy's Hospital, King's College, London, UK.
D000908:Antibodies, Fungal / Q000097:blood*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006031:Glycosylation; D006659:Histoplasmin / Q000276:immunology / Q000302:isolation & purification*; D006660:Histoplasmosis / Q000097:blood / Q000175:diagnosis*; D006801:Humans; D012680:Sensitivity and Specificity
D000908:Antibodies, Fungal; D006659:Histoplasmin