Evaluation of methodology for detection of human adenoviruses in wastewater, drinking water, stream water and recreational waters

J Appl Microbiol. 2010 Mar;108(3):800-809. doi: 10.1111/j.1365-2672.2009.04477.x. Epub 2009 Jul 13.

Abstract

Aims: This study evaluates dialysis filtration and a range of PCR detection methods for identification and quantification of human adenoviruses in a range of environmental waters.

Methods and results: Adenovirus was concentrated from large volumes (50-200 l) of environmental and potable water by hollow fibre microfiltration using commercial dialysis filters. By this method, an acceptable recovery of a seeded control bacteriophage MS2 from seawater (median 95.5%, range 36-98%, n=5), stream water (median 84.7%, range 23-94%, n=5) and storm water (median 59.5%, range 6.3-112%, n=5) was achieved. Adenovirus detection using integrated cell culture PCR (ICC-PCR), direct PCR, nested PCR, real-time quantitative PCR (qPCR) and adenovirus group F-specific direct PCR was tested with PCR products sequenced for confirmation. Adenovirus was routinely detected from all water types by most methods, with ICC-PCR more sensitive than direct-nested PCR or qPCR. Group F adenovirus dominated in wastewater samples but was detected very infrequently in environmental waters.

Conclusions and implications: Human adenoviruses (HAdv) proved relatively common in environmental and potable waters when assessed using an efficient concentration method and sensitive detection method. ICC-PCR proved most sensitive, could be used semiquantitatively and demonstrated virus infectivity but was time consuming and expensive. qPCR provided quantitative results but was c. ten-fold less sensitive than the best methods.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenoviruses, Human / isolation & purification*
  • Bathing Beaches
  • Cell Line
  • DNA, Viral / isolation & purification
  • Environmental Monitoring / methods*
  • Filtration / methods
  • Humans
  • Pilot Projects
  • Polymerase Chain Reaction / methods
  • Recreation
  • Rivers / virology*
  • Seawater / virology
  • Sensitivity and Specificity
  • Waste Disposal, Fluid
  • Water Microbiology*
  • Water Pollutants / isolation & purification
  • Water Pollution / analysis
  • Water Supply*

Substances

  • DNA, Viral
  • Water Pollutants