Detection of enteric viruses in municipal sewage sludge by a combination of the enzymatic virus elution method and RT-PCR

Daisuke Sano, Kensuke Fukushi, Yasuko Yoshida, Tatsuo Omura

Research output: Contribution to journalArticlepeer-review

29 Citations (Scopus)

Abstract

Pathogenic enteric viruses can be retained in municipal sewage sludge as has been reported by many researchers. Although the RT-PCR technique has been extensively employed for the virus detection from various environmental samples, the application of RT-PCR to the detection of viruses in sewage sludge has the difficulty because of inhibitory substances to the gene amplification. However, a combination of the enzymatic virus elution (EVE) method with RT-PCR made it possible to effectively detect viruses in sewage sludge. The enzymatic breakdown of sludge flocs in the EVE method enhanced the virus elution from poliovirus 1 (PV1)-inoculated sewage sludge, and the detection of PV1 was performed by RT-PCR without any inhibitions. On the contrary, the application of RT-PCR to the viral assay in the USEPA method using the 10% beef extract solution was not practical because of inhibitions to the viral gene amplification. The combination of the EVE method using lysozyme (polysaccharide-degrading enzyme), papain (protease), and chymotrypsin (protease) with RT-PCR resulted in a virus recovery efficiency of 31%, but a synergistic effect of these enzymes on the virus recovery efficiency was not observed. The EVE method using lysozyme or papain could be a promising procedure for the virus elution from sewage sludge in detecting these viruses with RT-PCR.

Original languageEnglish
Pages (from-to)3490-3498
Number of pages9
JournalWater Research
Volume37
Issue number14
DOIs
Publication statusPublished - 2003 Aug

Keywords

  • Cation exchange resin
  • Enzymatic virus elution method
  • Hydrolytic enzymes
  • Municipal sewage sludge
  • Pathogenic enteric viruses
  • RT-PCR

ASJC Scopus subject areas

  • Ecological Modelling
  • Water Science and Technology
  • Waste Management and Disposal
  • Pollution

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