RESUMO
We cloned the goose heat shock protein 70 gene (HSP70), to determine its sequence variation and elucidate its mRNA expression. We designed primers to amplify the entire goose HSP70 sequence. We used 10 commercial Wuzong goslings in a heat-stress experiment. We collected tissue samples for RNA extraction and quantitative real-time polymerase chain reaction (qRT-PCR). We analyzed the variation in expression of goose HSP70 before and after heat stress. We constructed a DNA pool from six different species, for single nucleotide polymorphism (SNP) screening. We detected 18 SNPs and selected three of these SNPs for correlation analysis with biological and immune traits in 200 Wuzong geese. We showed that T+237C was significantly correlated with the serum corticosterone level, whereas T+1122C was significantly correlated with the heterophil to lymphocyte ratio. Goose HSP70 contained no introns. The results of qRT-PCR analysis revealed significant gender differences in the expression of goose HSP70 at 40°C but not at 25°C; moreover, in general, expression was significantly higher at 40°C than at 25°C. With the exception of the leg muscle and cerebellum, HSP70 expression was significantly higher in male geese than in female geese. Our results indicate that goose HSP70 plays an important role in response to severe heat stress.
Assuntos
Gansos/genética , Gansos/imunologia , Regulação da Expressão Gênica , Proteínas de Choque Térmico HSP70/sangue , Proteínas de Choque Térmico HSP70/genética , Polimorfismo Genético/genética , Estresse Fisiológico , Animais , Feminino , Gansos/sangue , Proteínas de Choque Térmico HSP70/fisiologia , Masculino , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Estresse Fisiológico/genética , Estresse Fisiológico/imunologiaRESUMO
A new reagent was designed to the indirect hemagglutination test (IHATIAL), utilizing goose red blood cells as inert matrix and standardized for the field diagnosis of American trypanosomiasis. The objective was to substitute the lyophilized or frozen reagent of IHAT produced routinely using human erythrocytes in the Adolfo Lutz Institute (Säo Paulo/Brazil). The standardized reagent presented a long stability in liquid suspension, and was evaluated in 137 serum samples from patient with and without Chagas' disease, by IHATIAL. The diagnostic performance of this test was similar to the IHAT utilizing human erythrocytes and to that of a commercial IHAT kit. The sensitivity was 1.00, specificity 0.98, predictive value of positive 0.96 and of negative 1.00. Different batches of reagent successively produced proved to be reproducible in a quality control method. The new reagent is more economic than the former reagent, it can be produced easily and may be applicable to the seroepidemiologic studies.
Assuntos
Humanos , Animais , Gansos/imunologia , Doença de Chagas/diagnóstico , Testes de Hemaglutinação/métodos , Sensibilidade e Especificidade , Gansos/sangue , Indicadores e ReagentesRESUMO
A new reagent was designed to the indirect hemagglutination test (IHATIAL), utilizing goose red blood cells as inert matrix and standardized for the field diagnosis of American trypanosomiasis. The objective was to substitute the lyophilized or frozen reagent of IHAT produced routinely using human erythrocytes in the Adolfo Lutz Institute (São Paulo/Brazil). The standardized reagent presented a long stability in liquid suspension, and was evaluated in 137 serum samples from patient with and without Chagas' disease, by IHATIAL. The diagnostic performance of this test was similar to the IHAT utilizing human erythrocytes and to that of a commercial IHAT kit. The sensitivity was 1.00, specificity 0.98, predictive value of positive 0.96 and of negative 1.00. Different batches of reagent successively produced proved to be reproducible in a quality control method. The new reagent is more economic than the former reagent, it can be produced easily and may be applicable to the seroepidemiologic studies.