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A fluorescence‐based method coupled with Disruptor filtration for rapid detection of F + RNA phages
Author(s) -
Yang Y.,
Griffiths M. W.
Publication year - 2014
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/lam.12173
Subject(s) - rna , lytic cycle , filtration (mathematics) , fluorescence , biology , chromatography , bacteriophage , contamination , detection limit , microbiology and biotechnology , chemistry , virology , escherichia coli , biochemistry , gene , virus , physics , ecology , statistics , mathematics , quantum mechanics
Abstract 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.