Full Description

A rapid and simple immunomagnetic multiple capture reverse transcription-polymerasechain reaction method, (IMMC-RT-PCR), was developed to collect anddetect viruses listed on the US Environmental Protection Agency's (USEPA's) Contaminate Candidate List (CCL). In this proceduremonoclonal mouse anti-enterovirus and monoclonal mouse anti-adenovirus primaryantibodies, targeting the CCL viruses, was added to virus-spiked samples that had beenconcentrated using centrifugal filters. The concentrated samples were then purifiedusing sephadex G-200 spun columns. The virus antibody complex was separated fromthe solution using a secondary antibody attached to magnetic beads. The viral RNAwas then heat extracted and RT-PCR was performed. Detection limits for this methodwere 5 pfu for adenovirus serotypes 40, (AD40), and 41, (AD 41), 20 pfu for echovirus9, (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 theIMMC-RT-PCR method. The primers, however, were virus type specific and thedetection limits for ECV9 and CVA9 were unacceptable. Untreated influent samples oftwo local water treatment facilities, Alfred Merritt Smith, (AMSWTF), and RiverMountain, (RMWTF), were also collected following the ICR sample collection protocolsand then analyzed for enteric viruses using IMMC-RT-PCR. Includes 28 references, tables, figures.

Product Details

Edition: Vol. - No. Published: 11/01/2005 Number of Pages: 14File Size: 1 file , 1.3 MB