Abstract
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-RNA coliphage levels ≥3.2 log plaque forming units (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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References
Anon. (1995). ISO 10705-1. Water quality-detection and enumeration of bacteriophages. Part 1. Enumeration of F-specific RNA bacteriophages. Geneva: International Organization for Standardization.
Anon. (2001). USEPA Method 1602. Male specific and somatic coliphage in water by single agar layer (SAL) procedure. Washington, DC: United States Environmental Protection Agency, Office of Water.
Atmar, R. L., & Estes, M. K. (2001). Diagnosis of noncultivatable gastroenteritis viruses, the human caliciviruses. Clinical Microbiology Reviews, 14(1), 15–37.
Bidawid, S., Malik, N., Adegbunrin, O., Sattar, S. A., & Farber, J. M. (2003). A feline kidney cell line-based plaque assay for feline calicivirus, a surrogate for Norwalk virus. Journal of Virological Methods, 107(3), 163–167.
Bollback, J. P., & Huelsenbeck, J. P. (2001). Phylogeny, genome evolution, and host specificity of single stranded RNA bacteriophage (Family Leviviridae). Journal of Molecular Evolution, 52(2), 117–128.
Cirino, N. M., Tavakoli, N. P., Madison-Antenucci, S., & Egan, C. (2007). Multiplex rtPCR in microbiology. In I. M. Mackay (Ed.), Real-time PCR in microbiology: From diagnosis to characterization (pp. 183–229). Norfolk: Caister Academic Press.
Cole, D., Long, S. C., & Sobsey, M. D. (2003). Evaluation of F+RNA and DNA coliphages as source-specific indicators of fecal contamination in surface waters. Applied and Environmental Microbiology, 69(11), 6507–6514.
Cook, N., Bridger, J., Kendall, K., Gomara, M. I., El-Attar, L., & Gray, J. (2004). The zoonotic potential of rotavirus. Journal of Infection, 48(4), 289–302.
Dawson, D. J., Paish, A., Staffel, L. M., Seymour, I. J., & Appleton, H. (2005). Survival of viruses on fresh produce, using MS2 as a surrogate for norovirus. Journal of Applied Microbiology, 98(1), 203–209.
Doré, W. J., Henshilwood, K., & Lees, D. N. (2000). Evaluation of F-specific RNA bacteriophage as a candidate human enteric virus indicator for bivalve moluscan shellfish. Applied and Environmental Microbiology, 66(4), 1280–1285.
Farkas, T., Nakajima, S., Sugieda, M., Deng, X., Zhong, W., & Jiang, X. (2005). Seroprevalence of norovirus in swine. Journal of Clinical Microbiology, 43(2), 657–661.
Formiga-Cruz, M., Allard, A. K., Conden-Hansson, A. C., Henshilwood, K., Hernroth, B. E., Jofre, J., et al. (2003). Evaluation of potential indicators of viral contamination in shellfish and their applicability to diverse geographical areas. Applied and Environmental Microbiology, 69(3), 1556–1563.
Furuse, K. (1987). Distribution of coliphages in the environment: General considerations. In S. G. Goyal, C. P. Gerba, & G. Bitton (Eds.), Phage ecology (pp. 87–124). New York: Wiley.
Havelaar, A. H., Hogeboom, W. M., & Pot, R. (1984). F-specific RNA bacteriophages in sewage: Methodology and occurrence. Water Science and Technology, 17(4/5), 645–655.
Kirs, M., & Smith, D. C. (2007). Multiplex quantitative real-time reverse transcriptase PCR for F+-specific RNA coliphages: A method for use in microbial source tracking. Applied and Environmental Microbiology, 73(3), 808–814.
Mattison, K., Brassard, J., Houde, A., Simard, C., Pagotto, F., Jones, T., et al. (2007a). The feline calicivirus (FCV) as an internal control for the detection of RNA viruses from foods. Federal food safety and nutrition research meeting. Winnipeg, Manitoba, Canada.
Mattison, J., Shukla, A., Cook, A., Pollari, F., Friendship, R., Kelton, D., et al. (2007b). Human noroviruses in swine and cattle. Emerging Infectious Diseases, 13(8), 1184–1188.
Mead, P. S., Slutsker, L., Dietz, V., McCaig, L. F., Bresee, J. S., Shapiro, C., et al. (1999). Food-related illness and death in the United States. Emerging Infectious Diseases, 5(5), 607–625.
O’Connell, K. P., Bucher, J. R., Anderson, P. E., Cao, C. J., Khan, A. S., Gostomski, M. V., et al. (2006). Real-time fluorogenic reverse transcription-PCR assays for detection of bacteriophage MS2. Applied and Environmental Microbiology, 72(1), 478–483.
Ogorzaly, L., & Gantzer, C. (2006). Development of real-time RT-PCR methods for specific detection of F-specific RNA bacteriophage genogroups: Application to urban raw wastewater. Journal of Virological Methods, 138(1–2), 131–139.
Ogorzaly, L., & Gantzer, C. (2007). Erratum to “Development of real-time RT-PCR methods for specific detection of F-specific RNA bacteriophage genogroups: Application to urban raw wastewater”. J. Virol. Meth. 138 (2006):131–139. Journal of Virological Methods, 143(1), 122.
Schaper, M., Jofre, J., Uys, M., & Grabow, W. O. K. (2002). Distribution of genotypes of F-specific RNA bacteriophages in human and non-human sources of faecal pollution in South Africa and Spain. Journal of Applied Microbiology, 92(4), 657–667.
Scott, T. M., Rose, J. B., Jenkins, T. M., Farrah, S. R., & Lukasik, J. (2002). Microbial source tracking: Current methodology and future directions. Applied and Environmental Microbiology, 68(12), 5796–5803.
Sobsey, M. D., Battigelli, D. A., Shin, G. A., & Newland, S. (1998). RT-PCR amplification detects inactivated viruses in water and wastewater. Water Science and Technology, 38(12), 91–94.
Sundram, A., Jumanlal, N., & Ehlers, M. M. (2006). Genotyping of F-RNA coliphages isolated from wastewater and river water samples. Water SA, 32(1), 65–70.
Vinjé, J., Oudejans, S. J. G., Stewart, J. R., Sobsey, M. D., & Long, S. C. (2004). Molecular detection and genotyping of male-specific coliphages by reverse transcription-PCR and reverse line blot hybridization. Applied and Environmental Microbiology, 70(10), 5996–6004.
Wang, Q. H., Han, M. G., Cheetham, S., Souza, M., Funk, J. A., & Saif, L. J. (2005). Porcine noroviruses related to human noroviruses. Emerging Infectious Diseases, 11(12), 1874–1881.
Ward, P., Müller, P., Letellier, A., Quessy, S., Simard, C., Trottier, Y. L., et al. (2008). Molecular characterization of Hepatitis E virus detected in swine farms in the province of Quebec. Canadian Journal of Veterinary Research, 72(1), 27–31.
Wolf, S., Hewitt, J., Rivera-Aban, M., & Greening, G. E. (2008). Detection and characterization of F+ RNA bacteriophages in water and shellfish: Application of a multiplex real-time reverse transcription PCR. Journal of Virological Methods, 149(1), 123–128.
Acknowledgments
The authors thank Danielle Leblanc at AAFC, Food Research and Development Centre, St-Hyacinthe, QC, Canada, for her valuable discussion and FCV stock production and titration. This research was supported by Agriculture and Agri-Food Canada Research Branch Peer Reviewed Research Projects 75 and 162.
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Jones, T.H., Houde, A., Poitras, E. et al. 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. Food Environ Virol 1, 57–65 (2009). https://doi.org/10.1007/s12560-009-9008-7
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DOI: https://doi.org/10.1007/s12560-009-9008-7