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

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.