AWWA WQTC62555

A rapid and simple immunomagnetic multiple capture reverse transcription-polymerase chain reaction method, (IMMC-RT-PCR), was developed to collect and detect viruses listed on the US Environmental Protection Agency's (USEPA's) Contaminate Candidate List (CCL). In this procedure monoclonal mouse anti-enterovirus and monoclonal mouse anti-adenovirus primary antibodies, targeting the CCL viruses, was added to virus-spiked samples that had been concentrated using centrifugal filters. The concentrated samples were then purified using sephadex G-200 spun columns. The virus antibody complex was separated from the solution using a secondary antibody attached to magnetic beads. The viral RNA was then heat extracted and RT-PCR was performed. Detection limits for this method were 5 pfu for adenovirus serotypes 40, (AD40), and 41, (AD 41), 20 pfu for echovirus 9, (ECV9), 20 pfu for coxsackievirus A9, (CVA9), and 20 pfu for coxsackievirus B1, (CVB1). Designed primers that were specific for each virus group were tested using the IMMC-RT-PCR method. The primers, however, were virus type specific and the detection limits for ECV9 and CVA9 were unacceptable. Untreated influent samples of two local water treatment facilities, Alfred Merritt Smith, (AMSWTF), and River Mountain, (RMWTF), were also collected following the ICR sample collection protocols and then analyzed for enteric viruses using IMMC-RT-PCR. Includes 28 references, tables, figures.