Comparative efficacy of fluorescent antibody test, immunoperoxidase test and enzyme linked immuno sorbent assay in detection of rotavirus in cell culture

Virusdisease. 2014;25(2):239-42. doi: 10.1007/s13337-014-0196-x. Epub 2014 Feb 7.

Abstract

Rotavirus is prevalent worldwide and has been established as a leading cause of mortality due to severe diarrhoea in neonates. Isolation of the virus is a gold standard method for confirmation of rotavirus infection in the host. Propagation of rotavirus in cell culture is a challenge as in many instances the virus does not produce detectable cytopathic effect. This study was undertaken to evaluate the efficacy of fluorescent antibody test (FAT), immunoperoxidase test (IPT) and sandwich ELISA (S-ELISA) to detect rotavirus antigen propagated in MA 104 cell line. The intensity of fluorescence and colour development for I-FAT and I-IPT was categorized and the ELISA OD values were analyzed. The overall mean of detection were 5.16 ± 0.47, 5.16 ± 0.54 and 5.66 ± 0.33 for I-FAT, I-IPT and S-ELISA, respectively. Significantly less number of samples were positive in the initial one or two passage, which increased up to 100 % from third/fourth passage onwards. The study concluded that I-FAT, I-IPT and S-ELISA were equally effective in detecting propagated rotavirus in cell line, and the former two tests are suitable for in situ demonstration of the virus while the later could be used to assay antigen in cell culture fluid.

Keywords: Cell culture; ELISA; FAT; IPT; Rotavirus.