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.
| Edition : | Vol. - No. |
| File Size : | 1
file
, 1.3 MB |
| Note : | This product is unavailable in Ukraine, Russia, Belarus |
| Number of Pages : | 14 |
| Published : | 11/01/2005 |
