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Development and evaluation of a multiplexed real-time TaqMan RT-PCR assay with a sample process control for detection of F-specific RNA coliphage genogroups I and IV

Jones, T.H., Houde, A., Poitras, E., Ward, P., Johns, M.W. (2009). Development and evaluation of a multiplexed real-time TaqMan RT-PCR assay with a sample process control for detection of F-specific RNA coliphage genogroups I and IV, 1(2), 57-65. http://dx.doi.org/10.1007/s12560-009-9008-7

Abstract

© Her Majesty the Queen in Right of Canada 2009. There are increasing concerns of zoonotic transmission of some animal enteric viruses, such as calicivirus, hepatitis E virus, and rotavirus, which are closely related to human pathogenic strains. Most enteric viruses are detected by molecular techniques because they cannot be cultured. Surrogates such as F-RNA coliphages are cultivable but few molecular methods exist. Individual real-time TaqMan RT-PCR assays for the replicase gene of F-RNA coliphage genogroups I and IV were developed and multiplexed with a real-time TaqMan RT-PCR assay for feline calicivirus as a sample process control for the simultaneous detection and enumeration of genogroup I and IV F-RNA coliphages. Genogroup IV were successfully detected with the multiplexed assay in 80% of fecal samples that contained F-RNAcoliphagelevels ≥3.2 log plaque formingunits(pfu). F-RNA coliphage were at or below the limit of detection in most fecal samples when levels were ≤4 log pfu/g.

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