Title of article :
Detection of enteric viruses in municipal sewage sludge by a combination of the enzymatic virus elution method and RT-PCR
Author/Authors :
Sano، Daisuke نويسنده , , Fukushi، Kensuke نويسنده , , Yoshida، Yasuko نويسنده , , Omura، Tatsuo نويسنده ,
Issue Information :
روزنامه با شماره پیاپی سال 2003
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.
Keywords :
Municipal sewage sludge , Pathogenic enteric viruses , RT-PCR , Enzymatic virus elution method , Hydrolytic enzymes , Cation exchange resin
Journal title :
Water Research
Journal title :
Water Research