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Male-specific coliphages (FRNA) have been used as fecal indicators andpossible viral indicators for various water matrices such as surface, ground, anddrinking water. US Environmental Protection Method Method 1602 has been validated to detect andenumerate FRNA coliphages from groundwater. However, it requires 18-24 hrincubation period and may not be suitable for emerging detection of fecalcontamination in drinking water. Real-time PCR is one of the latest developedtechnologies that have been used in clinical microbiology to rapidly detect andquantify microorganisms successfully. It allows real time monitoring on theamplification process and quantifying the fluorescent labeled amplicons in eachcycle. The emitted fluorescent signal increases in direct proportion tothe amount of amplicons in a reaction. This study explores thepossibility of rapidly quantifying FRNA coliphages in river and raw water usingreal-time PCR. One mL raw water was spiked with a reference strain ofcoliphages MS2, followed by passing through size exclusion-ion exchangecolumn prior to real-time RT-PCR reaction. The chelax-100 and sephedex-100, size exclusion-ion exchange column was found to effectively remove PCRinhibitors in raw water because no coliphage MS2 was detected without treatedwith the chelax-sephedex column. The detection sensitivity of the real-time RTPCRdeveloped 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 waterand twelve raw water samples collected from the Semenyih Treatment Plant,Selangor over the period of 5 months, 40% of the samples were positive forFRNA with a range from 1 to 12 PFU/reaction by real-time RT-PCR, but wereenumerated with plaque counts ranged from 2 to 44 PFU/100ml by plaqueanalysis. Includes table, figures.

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Edition: Vol. - No. Published: 11/01/2005 Number of Pages: 12File Size: 1 file , 810 KB