Journal
JOURNAL OF VIROLOGICAL METHODS
Volume 149, Issue 1, Pages 123-128Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jviromet.2007.12.012
Keywords
F plus RNA bacteriophages; real-time RT-PCR; microbial source tracking
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Genotyping of F+ RNA bacteriophages has been used to distinguish between human and animal contributions to contaminated water and food. There are four genetically distinct genogroups of F+ RNA bacteriophages. Genogroups I and IV predominate in animal wastes and genogroups II and III in wastes of human origin. In this study, a multiplex real-time RT-PCR-based method was developed to detect and genotype F+ RNA bacteriophages. The assay was shown to be broadly reactive against a wide spectrum of F+ RNA bacteriophage strains, including MS2, GA, Q beta, MX1, SP and FI, and was able to detect and genotype F+ RNA bacteriophages in shellfish and river water. The assay is highly sensitive, with detection limits <10 PFU/reaction and <10 copies/reaction of the target sequences carried in plasmids, respectively. The applications of this assay include F+ RNA semi-quantitation and microbial source tracking. (C) 2008 Elsevier B.V. All rights reserved.
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