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J Virol Methods. 2008 Apr;149(1):123-8. doi: 10.1016/j.jviromet.2007.12.012. Epub 2008 Feb 15.

Detection and characterization of F+ RNA bacteriophages in water and shellfish: application of a multiplex real-time reverse transcription PCR.

Author information

1
Communicable Disease Group, Institute of Environmental Science & Research Ltd., Kenepuru Science Centre, PO Box 50-348, Porirua, New Zealand.

Abstract

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.

PMID:
18280588
DOI:
10.1016/j.jviromet.2007.12.012
[Indexed for MEDLINE]

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