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A fluorescence‐based method coupled with Disruptor filtration for rapid detection of F + RNA phages

Yang, Y., Griffiths, M. W.
Letters in applied microbiology 2014 v.58 no.2 pp. 177-183
RNA, bacteriophages, beta-galactosidase, enzyme activity, filtration, fresh produce, pollution
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⁻¹of F+RNA phages within 3 h, while 0·01 PFU ml⁻¹was 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.