Journal of Applied Microbiology, Vol.108, No.3, 800-809, 2010
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 center dot 5%, range 36-98%, n = 5), stream water (median 84 center dot 7%, range 23-94%, n = 5) and storm water (median 59 center dot 5%, range 6 center dot 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.
Keywords:adenovirus;drinking water;environmental water;integrated cell culture PCR;rapid techniques qPCR;water quality