ABSTRACT
There was developed a polyclonal “double-sandwich” antibody ELISA method for detection of rotavirus from stool specimens. The NCDV strain of rotavirus group A was concentrated by ultra-centrifugation using a 45% (v/v) saccharose cushion and inoculated in rabbits and sheep. 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 unspecific reactions. The presence of rotavirus was detected by the sheep’s IgG and revealed by the rabbit’s IgG, using an anti-rabbit IgG peroxidase conjugate developed in goats. When applied in population of 86 fecal samples from piglets with diarrhea, the results of the ELISA were: 100% of sensivity; 98.79% of specificity, with an agreement of 98.83%, using as standard the polyacrilamide gel electrophoresis (PAGE) test. The variances 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 alternative test for rotavirus diagnosis from fecal material.
KEY WORDS:
Rotavirus; diarrhea; PAGE; ELISA.