Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 2 de 2
Filtrar
1.
Artigo em Inglês | IMSEAR | ID: sea-21909

RESUMO

BACKGROUND & OBJECTIVES: Leptospirosis, a zoonosis with a worldwide distribution is an acute febrile illness caused by spirochaetes of the pathogenic Leptospira interrogans. Microscopic agglutination test (MAT), the reference method for diagnosis was successively done to evaluate the modified ELISA which was developed with the recombinant LipL32 antigen for the detection of anti-leptospiral antibodies in human serum samples. METHODS: The recombinant LipL32 antigen was developed from the serovar Pomona strain Pomona of the pathogenic L. interrogans species. The predicted titre at a single working dilution was plotted against the observed antiserum titre. Subsequently, predicted antibody activity titres were determined directly from the standard curve by solving the regression line equation. The relative sensitivity, specificity and accuracy of the single dilution ELISA for the detection of anti-leptospiral antibodies were determined in comparison to the MAT. RESULTS: A linear relationship was found between the predicted antibody titres at a single working dilution of 1:250 and the corresponding observed serum titres by the standard serial-dilution method. Regression analysis was used to determine a standard curve from which an equation was derived that allowed demonstration of the mentioned correlation. The equation was then used to convert the corrected absorbance readings of the single working dilution directly into the predicted ELISA antibody titres. A high level of sensitivity of 96 per cent and specificity of 91 per cent between ELISA and MAT titres was found. The kappa value was almost 1.0 indicating perfect agreement. INTERPRETATION & CONCLUSIONS: The r LipL32 ELISA was proved to be sensitive, specific and accurate as compared to the standard MAT and the test could be efficiently utilized as a screening test for a large number of human serum samples for the detection of leptospiral antibodies.


Assuntos
Antígenos de Bactérias/sangue , Proteínas da Membrana Bacteriana Externa/sangue , Ensaio de Imunoadsorção Enzimática/métodos , Humanos , Leptospira interrogans/imunologia , Leptospirose/diagnóstico , Lipoproteínas/sangue , Análise de Regressão , Sensibilidade e Especificidade , Titulação por Diluição de Reatividade a Testes Cutâneos
2.
Indian J Exp Biol ; 1997 Aug; 35(8): 903-5
Artigo em Inglês | IMSEAR | ID: sea-61277

RESUMO

Three different methods, namely guanidine hydrochloride, phenol-chloroform extraction and high salt method, were compared in order to develop a simple method for the extraction of high yield of DNA from buffalo blood suitable for genome analysis. Both phenol-chloroform and high salt methods produced good yields of high molecular weight DNA as determined by agarose gel electrophoresis. The yield and quality of DNA extracted by high salt method was comparable to that of phenol-chloroform method. The mean yields of DNA from 10 ml of whole blood extracted by either the phenol-chloroform or the high salt methods were 446.16 micrograms (SE = 26.68) and 432.83 micrograms (SE = 19.34) respectively. The DNA obtained from both methods was suitable for conventional as well as polymerase chain reaction (PCR) based restriction fragment length polymorphism (RFLP) studies. Extraction using the guanidine hydrochloride method resulted in a gelatinous material that failed to resuspend in TE buffer. The high salt method is quick and reliable and can be routinely used for the extraction of DNA from buffalo samples instead of phenol-chloroform extraction which is hazardous and time-consuming.


Assuntos
Animais , Búfalos/genética , DNA/isolamento & purificação , Genoma , Leucócitos/química , Cloreto de Sódio , Fatores de Tempo
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA