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GREGORI, F. Neonatal diarrhea: development and evaluation of a method of ELISA for rotavirus detection from fecal material. [Diarreia neonatal: desenvolvimento e avaliacao de um metodo de ELISA para a deteccao de rotavirus a partir de material fecal]. Sao Paulo, 1999. 113p. Dissertacao (Mestrado) Faculdade de Medicina Veterinaria e Zootecnia, Universidade de Sao Paulo. Rotaviruses have been identified worldwide as a major etiologic agent of acute nonbacterial diarrhea in the young of many species, including humans. In this investigation was developed and evaluated a “double-sandwich” antibody ELISA method for detection of rotavirus from stool specimens. For that, the NCDV strain of rotavirus group A was serially cultivated in MA-104 cell culture. The virus was concentrated by ultra-centrifugation and inoculated in rabbits and sheeps. After that, the IgG of serum samples of the animals was purified by ion-exchange chromatography and absorbed with whole serum of both animal species using a glutaraldehyde polymer, in order to eliminate inespecific reactions. The presence of rotavirus was detected by the sheep’s IgG and revelated by the rabbit’s IgG, using a anti-rabit IgG peroxidase conjugate developed in goat. The values of diluition of the components of the ELISA and the cut-off value were defined using 26 fecal samples (13 positive and 13 negative) of piglets. Following this procedure, the test was employed in a panel of 86 fecal samples from piglets with diarrhea, using as standard the polyacrilamide gel electrophoresis (PAGE) test. The results of the ELISA were: 100% of sensivity; 98.79% of specificity, with an agreement of 98.83%. The variance between 86 repetitions of the same sample were 0.001 (for one positive sample) and 0.0002 (for one negative sample). These results showed that this ELISA is a sensitive and specific screening test for rotavirus diagnosis from fecal material. Uniterms: Rotavirus; ELISA; Diagnostic; Diarrhea; PAGE. ABREVIATURAS E SIMBOLOS %= porcento μg= micrograma μL= microlitro oC= graus Celsius apud= citado por BCA= acido bicinconinico DEAE= dietilaminoetil et al.= e colaboradores ELISA= enzyme-linked immunosorbent assay g= aceleracao da gravidade terrestre (9,8 m/s) K= quilo M= molar mA= miliamper mg=miligrama mL= mililitro mM= milimolar N= normal nm= nanometro p/v= peso/volume pH= concentracao de hidrogenio ionico q.s.p.= quantidade suficiente para RNA= acido ribonucleico SDS= dodecilsulfa to de sodio TEMED= tetrametiletilenodiamina TRIS= hidroximetil-aminometano UV= ultra-violeta v/v= volume/volume Observacao: Em funcao do uso consagrado na literatura tecnica, algumas abreviaturas utilizadas seguem as iniciais da sua grafia no idioma ingles.

Diarréia neonatal: desenvolvimento e avaliação de um método de 'Elisa' para a detecção de rotavírus a partir de material fecal.