The detection of astrovirus in sludge biosolids using an integrated cell culture nested PCR technique

The work presented here demonstrates the utility of the integrated cell cul ture-reverse transcriptase-polymerase chain reaction( ICC-RT-PCR) coupled w ith nested PCR to detect human astroviruses and enteroviruses in sludge bio solids. Viruses were concentrated by beef extract elution and organic flocc ulation prior to analysis by a plaque assay and ICC-RT-PCR. Astroviruses we re detected in all but one sample and all of the samples were positive for enteroviruses. We have demonstrated the prevalence and frequency of astrovi rus in sludge and validated the ICC-RT-PCR/nested PCR technique as a useful tool to detect viruses in sludge.