Detection and Characterization of Orthoreovirus in Alberta's Environmental Waters

Tyler T. Kostiuk
2015
Reoviruses have been shown to infect a very broad range of mammalian sources and infections in humans have been associated with gastrointestinal and respiratory illnesses. While the presence of reovirus in several different water matrices and clinical samples has been reported, the clinical implications of reovirus in Alberta's water and clinical samples have not been studied because of the lack of an appropriate screening method. To improve reovirus detection, a novel reverse-transcription
more » ... time PCR (qRT-PCR) assay was designed, developed, and implemented. Two sets of novel qPCR primers and probes (targeting the M1 and L3 genetic fragments) were designed and found to have sensitivities of 5 genetic copies per qPCR for M1 and 50 genetic copies per qPCR for L1, respectively. Using the newly developed qRT-PCR assay, reoviruses were detected in wastewater (15/16) and environmental surface water (20/216) samples obtained from various sites in Alberta, while no reoviruses were observed in clinical fecal samples obtained from patients suffering from gastroenteritis. Phylogenetic trees produced from S4 gene fragment sequences indicate that a variety of S4 alleles are present in the environment in Alberta, which is consistent with data previously published regarding reoviruses in the environment. iii Acknowledgements I would like to make several important acknowledgements at the conclusion of my Master's degree.
doi:10.7939/r3v97zx40 fatcat:jmxugmgtlva6blnakjd3mus45u