RESUMO
Congenital adrenal hyperplasia (CAH) is an autosomal recessive disorder caused by the deficiency of one of the five enzymes involved in the biosynthesis of corticosteroids. The most common form of the disease is the lack of 21-hydroxylase which provokes an accumulation of high levels of 17α-hydroxyprogesterone (17-OHP), the main biochemical marker for illness detection. Given the significance of neonatal diagnosis for ensuring a timely treatment to patients suffering from CAH, newborn screening is worldwide performed for the determination of 17-OHP from dried blood spots on filter paper. The non-specificity of antisera employed in immunoassays and the cross-reaction with fetal adrenal hormones produce an overestimation in the 17-OHP quantification. Immunization of mice with 17-OHP-3-(O-carboxymethyl) oxime-bovine serum albumin led to the generation of 15 anti-17-OHP IgG1-and-IgG2b-secreting hybridomas. The 6E2G9 monoclonal antibody presents cross-reactivity values similar to those achieved by rabbit antibodies employed in the solid phase of UMELISA® 17-OH Progesterona Neonatal, assay for the newborn screening of CAH in Cuba. Additionally, the use of 6E2G9 in the evaluation of dried blood spots samples from newborns on filter paper showed a decrease in the mean 17-OHP levels, thus demonstrating it can replace the conventional rabbit antisera.
Assuntos
17-alfa-Hidroxiprogesterona/sangue , Hiperplasia Suprarrenal Congênita/sangue , Anticorpos Monoclonais/sangue , Teste em Amostras de Sangue Seco , Ensaio de Imunoadsorção Enzimática , Triagem Neonatal , 17-alfa-Hidroxiprogesterona/imunologia , Hiperplasia Suprarrenal Congênita/diagnóstico , Animais , Anticorpos Monoclonais/imunologia , Biomarcadores/sangue , Estudos Transversais , Humanos , Recém-Nascido , Masculino , Camundongos , Camundongos Endogâmicos BALB CRESUMO
INTRODUCTION: The HIV-2 glycoprotein 36 (gp36) is often used in ELISA. An evaluation of the diagnostic indexes of antigen mixtures with a synthetic peptide of HIV2 gp36 (5) is performed in this study. METHODS: Five mixtures of gp36 (5) and the recombinant proteins of HIV1/2 were prepared. A total of 1306 samples were evaluated with UMELISA HIV1+2 RECOMBINANT used as reference. RESULTS: The variant (V-1) showed very good agreement as regards the reference method. CONCLUSION: The V-1 variant was shown to be highly effective in the immunodiagnosis of HIV 1/2.
Assuntos
Sorodiagnóstico da AIDS/métodos , Ensaio de Imunoadsorção Enzimática/métodos , Antígenos HIV/imunologia , Infecções por HIV/diagnóstico , HIV-1/imunologia , HIV-2/imunologia , Produtos do Gene env do Vírus da Imunodeficiência Humana/imunologia , Especificidade de Anticorpos , Anticorpos Anti-HIV/sangue , Anticorpos Anti-HIV/imunologia , Infecções por HIV/virologia , HIV-1/isolamento & purificação , HIV-2/isolamento & purificação , Humanos , Fragmentos de Peptídeos/síntese química , Fragmentos de Peptídeos/imunologia , Valor Preditivo dos Testes , Proteínas Recombinantes/síntese química , Proteínas Recombinantes/imunologia , Sensibilidade e Especificidade , Produtos do Gene env do Vírus da Imunodeficiência Humana/síntese químicaRESUMO
Six chimeric synthetic peptides (QCha-1, QCha-2, QCha-3, QCha-4, QCha-5, and QCha-6) incorporating antigenic sequences of two immunodominant repeat B-cell epitopes of Trypanosoma cruzi were synthesized by conventional solid-phase peptide synthesis. The antigenic activity of these peptides was evaluated by UltramicroEnzyme-linked immunosorbent assay (UMELISA) by using panels of positive Chagasic sera (n=82), while specificity was evaluated with samples from healthy blood donors (n=44) and patients with other infectious diseases (n=86). The antigenicity of the chimeric peptides in solid-phase immunoassays was compared with that of the monomeric peptides. Data demonstrated that the chimeric peptide QCha-5 was the most reactive because it detected antibodies to parasite efficiently. The results indicate that chimeric peptide as coating antigen is very useful for the immunodiagnosis of Chagas' disease.
Assuntos
Anticorpos Antiprotozoários/sangue , Doença de Chagas/diagnóstico , Epitopos de Linfócito B/imunologia , Peptídeos/imunologia , Trypanosoma cruzi/imunologia , Animais , Anticorpos Antiprotozoários/imunologia , Linfócitos B/imunologia , Linfócitos B/parasitologia , Doença de Chagas/imunologia , Doença de Chagas/parasitologia , Ensaio de Imunoadsorção Enzimática , Humanos , Peptídeos/síntese químicaRESUMO
A chimeric synthetic peptide incorporating immunodominant epitope of the p19 gag protein (116-134) and the gp46 env protein (178-200) of HTLV-II virus, separated by two glycine residues, was synthesized by conventional solid-phase peptide synthesis. The antigenic activity of this peptide was evaluated by Ultramicro Enzyme-linked immunosorbent assay (UMELISA) by using panels of anti-HTLV-II positive sera (n = 9), anti-HTLV-I/II positive sera (n = 2), HTLV-positive (untypeable) serum samples (n = 1),and anti-HTLV-I positive sera (n = 14), while specificity was evaluated with samples from healthy blood donors (n = 20). The efficacy of the chimeric peptide in solid-phase immunoassays was compared with the monomeric peptides. Data demonstrated that the chimeric peptide was the most reactive because it detected antibodies to virus efficiently. This may be related to peptide adsorption to the solid surface and epitope accessibility to the antibodies. The results indicate that chimeric peptide as coating antigen is very useful for the immunodiagnosis of HTLV-II infection.