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Updated: May 7, 2026
Detection of Viral RNA by Fluorescence in situ Hybridization FISH
Published on: May 5, 2012
A fluorescence-based method coupled with Disruptor filtration for rapid detection of F + RNA phages
1CRIFS, Department of Food Science, University of Guelph, Guelph, ON, Canada.
Unlabelled:
F + RNA phages are commonly used as indicators of faecal contamination. This study evaluated a fluorescent method for the detection of F + RNA phages based on testing the phage-mediated release of β-galactosidase. Factors that may potentially interfere with phage detection were investigated, and the assay was optimized. Low numbers of F + RNA phages were detected by the fluorescent method coupled with a concentration step using a Disruptor filter. The fluorescent method, when used alone, detected 1 log PFU ml(-1) of F+RNA phages within 3 h, while 0.01 PFU ml(-1) was detected within 5 h when the method was combined with the concentration step. This is the first time to combine a fluorescent method with a filtration step by the use of Disruptor filter for rapid detection of low numbers of F + RNA phages, and this method can be adapted to detect other lytic phages infecting host cells that produce measurable enzyme activity.
Significance And Impact Of The Study:
A fluorescent method coupled with Disruptor filtration was evaluated for the first time to rapidly detect low numbers of F + RNA phages. Rapid detection of F + RNA phages provides an effective way to monitor faecal contamination of environmental water and thus helps prevent contamination of fresh produce via irrigation.
Insights
A new fluorescent method combined with Disruptor filtration rapidly detects low levels of F+RNA phages. This advancement offers a faster way to monitor fecal contamination in water, safeguarding produce from contaminated irrigation.
Area of Science:
- Environmental microbiology
- Water quality monitoring
- Bacteriophage detection
Background:
- F+RNA phages are key indicators of fecal contamination in water sources.
- Accurate and rapid detection methods are crucial for public health and food safety.
- Existing methods for phage detection can be time-consuming.
Purpose of the Study:
- To evaluate a novel fluorescent method for F+RNA phage detection.
- To optimize the fluorescent assay for enhanced sensitivity and speed.
- To assess the efficacy of combining fluorescent detection with a concentration step.
Main Methods:
- Utilized a fluorescent assay measuring phage-mediated release of β-galactosidase.
- Investigated and optimized factors influencing phage detection.
- Employed a Disruptor filter for concentrating low phage numbers.
- Compared detection limits and times of the fluorescent method alone versus combined with filtration.
Main Results:
- The fluorescent method detected 1 log PFU/ml of F+RNA phages within 3 hours.
- Combining the fluorescent method with Disruptor filtration detected 0.01 PFU/ml within 5 hours.
- The optimized method demonstrated high sensitivity for detecting low phage concentrations.
Conclusions:
- The fluorescent method coupled with Disruptor filtration provides a rapid and sensitive approach for F+RNA phage detection.
- This method is the first to combine fluorescent detection with Disruptor filtration for rapid analysis of low phage numbers.
- The technique can be adapted for detecting other lytic phages and has implications for monitoring water quality and food safety.
