Strongyloides ratti antigenic components recognized by IgE antibodies in immunoblotting as an additional tool for improving the immunodiagnosis in human strongyloidiasis

IgE antibody response in human strongyloidiasis was evaluated by enzyme-linked immunosorbent assay (ELISA) and immunoblotting (IB) using Strongyloides ratti saline extract as heterologous antigen. A total of 50 serum samples of patients who were shedding S. stercoralis larvae in feces (group I, copropositive), 38 of patients with other intestinal parasites (group II), and 38 of subjects with negative results in three parasitologic assays (group III, copronegative) were analyzed. Levels of IgE anti-Strongyloides expressed in ELISA Index (EI) were significantly higher in patients of group I (1.32) than in group II (0.51) and group III (0.81), with positivity rates of 54 percent, 0 percent, and 10.5 percent, respectively. Fifteen S. ratti antigenic components were recognized in IB-IgE by sera of group I, with frequency ranging from 8 percent to 46 percent. In group II, only two antigenic bands (101, 81 kDa) were detected in a frequency of 10 percent and no reactivity was found in group III. Sera with EI values > 1.5 recognized five from 13 specific antigenic bands (70, 63, 61, 44, 7 kDa). It can be concluded that these five antigenic components recognized by IB-IgE using S. ratti antigen might be employed as an additional tool for improving the immunodiagnosis in human strongyloidiasis.

Heterologous antigen extract in ELISA for the detection of human IgE anti-Strongyloides stercoralis

Extrato contendo larvas de Strongyloides ratti foi usado na padronização de um ELISA para detecção de IgE gênero-específica na estrongiloidíase humana. Foram analisadas 40 amostras de soro de pacientes monoinfectados que estavam eliminando larvas de S. stercoralis nas fezes (Grupo I), 40 de pacientes com outros parasitos intestinais (Grupo II), e 40 indivíduos copronegativos (Grupo III). Níveis de IgE gênero-específica (índice ELISA: EI) foram significativamente maiores no Grupo I (EI = 1,43) do que no II (EI = 0,70) e III (EI = 0,71), mostrando positividade de 55 por cento, 2,5 por cento e 0 por cento, respectivamente. Similarmente, soros dos pacientes copropositivos (Grupo I) apresentaram níveis significativamente maiores de IgE total (866 IU/mL) quando comparados com os soros dos Grupo II (302 IU/mL) e III (143 IU/mL). Uma significativa correlação positiva foi encontrada entre os níveis de IgE específica a Strongyloides sp. e IgE total nos soros de pacientes com estrongiloidíase. Em conclusão, extrato heterólogo de S. ratti mostrou ser uma ferramenta útil para detecção de IgE gênero-específica por ELISA, desta forma contribuindo para melhor caracterização do perfil da resposta imune na estrongiloidíase humana.

Western blotting using Strongyloides ratti antigen for the detection of IgG antibodies as confirmatory test in human strongyloidiasis

The present study was conducted to evaluate the frequency of antigenic components recognized by serum IgG antibodies in Western blotting (WB) using a Strongyloides ratti larval extract for the diagnosis of human strongyloidiasis. In addition, the WB results were compared to the enzyme-linked immunosorbent assay (ELISA) and the indirect immunofluorescence antibody test (IFAT) results. Serum samples of 180 individuals were analyzed (80 with strongyloidiasis, 60 with other intestinal parasitoses, and 40 healthy individuals). S. ratti was obtained from fecal culture of experimentally infected Rattus rattus. For IFAT, S. ratti larvae were used as antigen and S. ratti larval antigenic extracts were employed in WB and ELISA. Eleven S. ratti antigenic components were predominantly recognized by IgG antibodies in sera of patients with strongyloidiasis. There was a positive concordance for the three tests in 87.5 percent of the cases of strongyloidiasis. The negative concordance in the three tests was 94 percent and 97.5 percent, in patients with other intestinal parasitoses and healthy individuals, respectively. In cases of positive ELISA and negative IFAT results, diagnosis could be confirmed by WB. ELISA, IFAT, and WB using S. ratti antigens showed a high rate of sensitivity and specificity. In conclusion, WB using S. ratti larval extract was able to recognize 11 immunodominant antigenic components, showing to be a useful tool to define the diagnosis in cases of equivocal serology