ABSTRACT
The aim of the study was to detect the rDNA sequences and their regions in Histoplasma capsulatum, which could be considered species-specific and used as a molecular method for this diagnosis by the technique of nested polymerase chain reaction (nested PCR), employing specific sequences (primers) for H. capsulatum: 18S rDNA region (HC18), 100 kDa (HC100) and the sequence 5.8 S-ITS rDNA (HC5.8). The PCR sequences HC18, HC100 and HC5.8 resulted in a specificity of 100%. The molecular assays may increase the specificity, sensitivity and speed in the diagnosis of Histoplasmosis.
Subject(s)
HIV Seropositivity/blood , Histoplasma/genetics , Histoplasmosis/diagnosis , Polymerase Chain Reaction , Early Diagnosis , Histoplasma/isolation & purification , Histoplasmosis/blood , Histoplasmosis/microbiology , Humans , Sensitivity and SpecificityABSTRACT
The aim of the study was to detect the rDNA sequences and their regions in Histoplasma capsulatum, which could be considered species-specific and used as a molecular method for this diagnosis by the technique of nested polymerase chain reaction (nested PCR), employing specific sequences (primers) for H. capsulatum: 18S rDNA region (HC18), 100 kDa (HC100) and the sequence 5.8 S-ITS rDNA (HC5.8). The PCR sequences HC18, HC100 and HC5.8 resulted in a specificity of 100%. The molecular assays may increase the specificity, sensitivity and speed in the diagnosis of Histoplasmosis.
O objetivo do estudo consistiu em detectar seqüências no ADNr e as suas regiões no Histoplasma capsulatum, que pudessem ser consideradas espécie-específicas e usadas como método molecular para o diagnóstico pela técnica da reação em cadeia da polimerase aninhada ("nested PCR") com seqüências específicas ("primers") para H. capsulatum: regiões 18S ADNr (HC18), 100kDa (HC100) e a seqüência 5.8 S ADNr-ITS (HC5.8). A "nested PCR" com as seqüências HC18, HC100 e HC5.8 resultaram em 100% de especificidade. Os ensaios moleculares podem aumentar a especificidade, sensibilidade e rapidez na diagnose da Histoplasmose.
Subject(s)
Humans , HIV Seropositivity/blood , Histoplasma/genetics , Histoplasmosis/diagnosis , Polymerase Chain Reaction , Early Diagnosis , Histoplasma/isolation & purification , Histoplasmosis/blood , Histoplasmosis/microbiology , Sensitivity and SpecificityABSTRACT
This study evaluated serum protein fractions, HDL-cholesterol, total immunoglobulin G and total immunoglobulin E levels in patients with acute and chronic paracoccidioidomycosis, by means of electrophoresis, enzymatic reaction and immunoenzymatic assay. The results demonstrated elevated levels of total immunoglobulin G, total immunoglobulin E, alpha-2 and gamma-globulins, which were more evident in acute than in chronic PCM, but no increase in HDL-cholesterol levels. There was a correlation between the levels of total immunoglobulin E and gamma-globulins and the alpha-2 and beta-globulin fractions in the acute form and between beta and gamma-globulins in both the acute and the chronic form. In conclusion, changes in total immunoglobulin G and immunoglobulin E levels and in the electrophoretic profile may be important markers for the prognosis and therapeutic follow-up of PCM cases, especially because protein electrophoresis is a simple laboratory test that can be applied when specific PCM serological tests are not available. In addition, levels of the gamma-globulin fraction greater than 2.0 g/dl may suggest that the patient is developing a more severe form of PCM.
Subject(s)
Blood Proteins/analysis , Cholesterol, HDL/blood , Immunoglobulin G/blood , Paracoccidioidomycosis/blood , Acute Disease , Adolescent , Adult , Aged , Aged, 80 and over , Biomarkers/blood , Child , Chronic Disease , Electrophoresis, Cellulose Acetate , Enzyme-Linked Immunosorbent Assay , Female , Humans , Immunodiffusion , Immunoglobulin E/blood , Male , Middle Aged , Paracoccidioidomycosis/immunology , Reference Values , Young AdultABSTRACT
This study evaluated serum protein fractions, HDL-cholesterol, total immunoglobulin G and total immunoglobulin E levels in patients with acute and chronic paracoccidioidomycosis, by means of electrophoresis, enzymatic reaction and immunoenzymatic assay. The results demonstrated elevated levels of total immunoglobulin G, total immunoglobulin E, alpha-2 and gamma-globulins, which were more evident in acute than in chronic PCM, but no increase in HDL-cholesterol levels. There was a correlation between the levels of total immunoglobulin E and gamma-globulins and the alpha-2 and beta-globulin fractions in the acute form and between beta and gamma-globulins in both the acute and the chronic form. In conclusion, changes in total immunoglobulin G and immunoglobulin E levels and in the electrophoretic profile may be important markers for the prognosis and therapeutic follow-up of PCM cases, especially because protein electrophoresis is a simple laboratory test that can be applied when specific PCM serological tests are not available. In addition, levels of the gamma-globulin fraction greater than 2.0g/dl may suggest that the patient is developing a more severe form of PCM.
Este trabalho avaliou as frações de proteínas séricas, HDL-colesterol e imunoglobulina G e imunoglobulina E totais em pacientes com paracoccidioidomicose aguda e crônica por eletroforese, reação enzimática e ensaio imunoenzimático. Os resultados demonstraram aumento dos níveis de imunoglobulina G e imunoglobulina E totais, alfa-2 e gama-globulinas, mais evidente na forma aguda que na forma crônica, e não nos níveis de HDL-colesterol. Houve correlação entre níveis de imunoglobulina E total e gama-globulinas e fração alfa-2 e beta-globulinas na forma aguda e entre beta e gama-globulinas nas formas aguda e crônica. Concluindo, alterações nos níveis de imunoglobulina G e imunoglobulina E totais e no perfil eletroforético podem ser importantes marcadores para prognóstico e acompanhamento terapêutico da PCM, especialmente por ser a eletroforese de proteínas um exame laboratorial simples que pode ser empregado em situações onde a sorologia específica para PCM não está disponível. Adicionalmente, níveis da fração gama-globulinas acima de 2,0g/dL podem sugerir que o paciente esteja desenvolvendo uma forma mais grave de PCM.
Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Child , Female , Humans , Male , Middle Aged , Young Adult , Blood Proteins/analysis , Cholesterol, HDL/blood , Immunoglobulin G/blood , Paracoccidioidomycosis/blood , Acute Disease , Biomarkers/blood , Chronic Disease , Electrophoresis, Cellulose Acetate , Enzyme-Linked Immunosorbent Assay , Immunodiffusion , Immunoglobulin E/blood , Paracoccidioidomycosis/immunology , Reference Values , Young AdultABSTRACT
No centésimo aniversário da publicação do primeiro caso de paracoccidioidomicose, são apresentadas informações relacionadas ao histórico, agente etiológico, epidemiologia, patogênese, formas clínicas, diagnóstico laboratorial e antígenos. Apesar do expressivo avanço em diversas áreas do conhecimento, a micose descrita por Adolpho Lutz em 1908 ainda demonstra altas taxas de mortalidade e letalidade e baixa visibilidade, configurando-se como oitava causa de mortalidade por doença predominantemente crônica ou repetitiva entre as infecciosas e parasitárias, e a maior taxa de mortalidade entre as micoses sistêmicas.
