Date of Award
Program or Major
Master of Science
Aaron B Margolin
The public health threat from pathogens creates controversy for the land application of biosolids, a sewage treatment byproduct. Previous work has demonstrated that some enteric viruses are not detected with a plaque assay, the current method for virus detection in biosolids. The Integrated Cell Culture - Quantitative Polymerase Chain Reaction (ICC-qPCR) assay, which combined quantitative PCR with seven days incubation in cell culture, allows for detection of more viruses.
To compare method sensitivities, a biosolid sample was seeded with mammalian orthoreovirus. 3x105 plaque forming units (PFU) per ml were detected by the plaque assay and 108 PFU equivalents per ml were detected by ICC-qPCR. To determine the ability of ICC-qPCR to detect mammalian orthoreovirus, twenty-four environmental samples were tested. No viruses were detected by the plaque assay based on the EPA method; however ICC-qPCR detected infectious mammalian orthoreovirus in thirteen samples. ICC-qPCR was more sensitive than the plaque assay.
Gallagher, Elizabeth, "Frequency of reovirus detection in biosolids: Comparison of the EPA CFR 503 technique to Integrated Cell Culture - Quantitative PCR" (2009). Master's Theses and Capstones. 529.