Male-specific coliphages (FRNA) have been used as fecal indicators and
possible viral indicators for various water matrices such as surface, ground, and
drinking water. US Environmental Protection Method Method 1602 has been validated to detect and
enumerate FRNA coliphages from groundwater. However, it requires 18-24 hr
incubation period and may not be suitable for emerging detection of fecal
contamination in drinking water. Real-time PCR is one of the latest developed
technologies that have been used in clinical microbiology to rapidly detect and
quantify microorganisms successfully. It allows real time monitoring on the
amplification process and quantifying the fluorescent labeled amplicons in each
cycle. The emitted fluorescent signal increases in direct proportion to
the amount of amplicons in a reaction. This study explores the
possibility of rapidly quantifying FRNA coliphages in river and raw water using
real-time PCR. One mL raw water was spiked with a reference strain of
coliphages MS2, followed by passing through size exclusion-ion exchange
column prior to real-time RT-PCR reaction. The chelax-100 and sephedex-100, size exclusion-ion exchange column was found to effectively remove PCR
inhibitors in raw water because no coliphage MS2 was detected without treated
with the chelax-sephedex column. The detection sensitivity of the real-time RTPCR
developed was 10 PFU per reaction. The primers were specific to detect all strains of FRNA but not FDNA and somatic coliphages. Of the eight river water
and twelve raw water samples collected from the Semenyih Treatment Plant,
Selangor over the period of 5 months, 40% of the samples were positive for
FRNA with a range from 1 to 12 PFU/reaction by real-time RT-PCR, but were
enumerated with plaque counts ranged from 2 to 44 PFU/100ml by plaque
analysis. Includes table, figures.
| Edition : | Vol. - No. |
| File Size : | 1
file
, 810 KB |
| Note : | This product is unavailable in Ukraine, Russia, Belarus |
| Number of Pages : | 12 |
| Published : | 11/01/2005 |
