Comparison of methods for rotavirus detection in water and results of a survey of Jerusalem wastewater

Abstract

Methods for the detection of viable rotaviruses and rotavirus antigen in water were developed and compared. The methods included laboratory-developed enzyme-linked immunosorbent assays (ELISAs) with chromogenic and luminescent substrates, commercial Rotazyme and Enzygnost ELISAs, and an indirect immunofluorescent assay. Of the methods tested, the immunofluorescent assay and the Enzygnost ELISA were the most sensitive for the simian rotavirus SA-li. All of the methods were positive for human rotavirus from clinical specimens. Seeded SA-li rotavirus was concentrated from water by adsorption to and elution from Zeta Plus ifiters followed by organic flocculation. Interference with the assays by components of the wastewater concentrates was minimal for the ELISAs, although the undiluted organic flocs were cytotoxic for the immunofluorescent assay. A survey of Jerusalem wastewater was carried out over the course of 1 year, and samples were assayed for rotaviruses and enteroviruses. Although enteroviruses were found in almost all of the samples, all samples were negative for rotaviruses. The concentration of rotaviruses in the wastewater was thus below the detection limit of the methods used.

Enteric viruses in the aquatic environment

“…Environmental samples have not often been tested directly by ELISA for rotavirus as insuf®cient VP6 antigen is present even after concentration (Guttmann-Bass et al 1987) and VP6-directed ELISA only detects noninfectious particles. Steinmann (1981) published the ®rst report of the identi®cation by ELISA of rotaviruses in 6/24 samples of sewage and Dahling et al (1993) reported rotavirus in three out of 30 sewage samples from Puerto Rico that had been concentrated by positive membrane ®ltration and were found to be positive by direct ELISA.…”

“…Environmental samples have not often been tested directly by ELISA for rotavirus as insufficient VP6 antigen is present even after concentration (Guttmann‐Bass et al . 1987) and VP6‐directed ELISA only detects noninfectious particles.…”