Multiparametric assay of screening for the diagnosis of mycoses of interest in Public Health: standardization of methodology

The standardization and validation of a multiplex assay requires the combination important parameters such as sensitivity specificity, acceptable levels performance, robustness, reproducibility. We standardized multiparametric Dot-blot aimed at serological screening paracoccidioidomycosis, histoplasmosis, aspergillosis. A total 148 serum were evaluated: 10 from healthy subjects, 36 patients with 62 40 It was found that showed high percentage cross-reactivity. However, when evaluated individually, in good performance observed compared to double immunodiffusion assay, considered gold standard test, 100% co-positivity 83.3% co-negativity. for aspergillosis not satisfactory immunodiffusion, showing 71.4% evaluation stability nitrocellulose membranes sensitized H. capsulatum antigen remained stable 90 days those A. fumigatus 30 days. conclude use crude antigens suitable due cross-reactivity, further tests should be performed purified proteins.