Assuntos
Meios de Cultura , Vaginite por Trichomonas/diagnóstico , Técnicas de Cultura de Células/métodos , Técnicas de Cultura de Células/normas , Feminino , Humanos , Prevalência , Fatores de Risco , Sensibilidade e Especificidade , Vaginite por Trichomonas/epidemiologia , Vaginite por Trichomonas/parasitologiaRESUMO
A simultaneous determination of retinol, alpha-tocopherol and beta-carotene in serum by high-performance liquid chromatography is described. Total analysis time is 13 min. A reversed-phase (Ultrasphere ODS, 5 microns) column is used with a mobile phase of acetonitrile-methanol-dichloromethane (70:10:20, v/v/v) and a flow-rate of 1.2 ml/min. Retinol is monitored at 325 nm, alpha-tocopherol at 292 nm and beta-carotene at 450 nm. Serum is deproteinized with ethanol containing the internal standard (alpha-tocopherol acetate), then extracted with hexane. The evaporated organic layer is reconstituted with the mobile phase and injected. The choice of the eluent is discussed, as well as the choice of an internal standard and the need for an antioxidant during the extraction step. Sixteen different eluents are compared in terms of analysis time and selectivity. The linear concentration ranges (retinol 0.016-13.7 microM, alpha-tocopherol 0.18-91.8 microM, beta-carotene 0.05-5.75 microM), within-run coefficients of variation (retinol less than 7%; alpha-tocopherol less than 8%, beta-carotene less than 7%), between-run coefficients of variation (retinol less than 13%, alpha-tocopherol less than 9%, beta-carotene less than 8%) and recoveries (retinol greater than 95%, alpha-tocopherol greater than 91%, beta-carotene greater than 80%) are suitable for clinical investigations. Serum reference values were found to be 2.47 +/- 0.61 microM (retinol), 30.5 +/- 6.8 microM (alpha-tocopherol) and 0.91 +/- 0.55 microM (beta-carotene). A significant difference (p less than 0.001) between males and females was found for retinol.
Assuntos
Carotenoides/sangue , Vitamina A/sangue , Vitamina E/sangue , Cromatografia Líquida de Alta Pressão , Feminino , Humanos , Masculino , Padrões de Referência , Valores de Referência , Reprodutibilidade dos Testes , Espectrofotometria Ultravioleta , beta CarotenoRESUMO
Aspergillus fumigatus antigens have been tested to determine their potential as aids in the diagnosis of invasive aspergillosis (IA). Immunoglobulin G (IgG) antibodies to these antigens were detected by analytical isoelectrofocusing in conjunction with immunoblotting. A total of 12 antigenic fractions, including culture filtrates and surface and mycelial extracts of A. fumigatus, were investigated. Eleven were reactive with serum specimens from patients with aspergilloma, which served as positive controls for the evaluation of a specific IgG response. Eight of 12 antigens showed good responses with serum specimens from patients with allergic bronchopulmonary aspergillosis, which were used to assess the sensitivity of IgG detection. No measurable reactivity was detected in 18 negative control serum specimens, while 11 of 13 patients with proven, highly probable, or probable cases of IA had anti-Aspergillus IgG to multiple antigenic preparations. Patients with IA who were capable of mounting a substantial humoral response to Aspergillus antigens gave an antibody profile with five antigenic preparations which seemed to be characteristic of the disease. Data show that this method is highly sensitive and may allow the selection of fractions which are both highly antigenic and specific for the detection of antibodies to Aspergillus antigens. They also indicate that the use of a spectrum of antigenic molecules is advisable, given the variability observed in the immune responses of individual patients.