PCR detection of specific pathogens in water: A risk-based analysis

The relative concentration of pathogens in water samples collected from storm drains and adjacent surfaces was evaluated using established PCR-based protocols, Out of the 58 samples collected from 21 different storm drains, 22% were PCR positive for Escherichia coli ETEC, Salmonella, or adenovirus, The risk of swimming related illnesses associated with detection of E coli ETEC and Salmonella ranged from 0.39 to 30:100 000 and 0.3-25:1000, respectively. The detection limit corresponding to a negative-PCR result was evaluated in reference to water quality standards developed using a risk-based approach that integrates human dose-response data with acceptable levels of risk promulgated by the U.S. EPA for recreational contact. The percent of samples with an acceptable detection limit ranged from 0% for Giardia lamblia and Shigella to 100% for E. coli ETEC. The principal factor influencing the detection limit of G. lamblia and Shigella was sample volume. The principal factor influencing the detection limit of the remaining bacteria and protozoa, including E coli O157:H7, Salmonella, and Cryptosporidium parvum, was the presence of inhibitory compounds in the purified nucleic acid extracts. Both recovery and inhibition adversely impacted the detection limit of viruses. Ambient water quality standards based on the occurrence of specific pathogens enumerated with PCR-based assays could serve as a method of evaluating the biological quality of water but only after significant improvements in filtration and purification protocols. The risk-based methodology developed in this study can be used to evaluate future improvements in filtration and purification protocols.