Development of a real time immuno-PCR assay for the detection of pea root rot causal agent, Aphanomyces euteiches

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dc.contributor.authorKaphle, Salina
dc.contributor.authorUniversity of Lethbridge. Faculty of Arts and Science
dc.contributor.supervisorYevtushenko, Dmytro
dc.date.accessioned2020-09-11T15:04:41Z
dc.date.available2020-09-11T15:04:41Z
dc.date.issued2020
dc.degree.levelMastersen_US
dc.description.abstractEffective measures to detect Aphanomyces euteiches, one of the devastating pathogen of pea, are lacking. An indirect ELISA and a real time immuno-PCR (RT-iPCR) assay for the timely and sensitive detection of A. euteiches were developed using antiserum specific to oospores of A. euteiches isolates. RT-iPCR was 100 times more sensitive over the linear working range than indirect ELISA. To assess the performance of the RT-iPCR assay, non-infested soils representing a range of soil textures were collected from Alberta fields. An extraction protocol was developed by spiking the soils with A. euteiches oospores and the RT-iPCR assay was used to quantify oospores in the soil extracts. The method performed well at the concentration 100 oospores/g and above. A rolled towel bioassay and root PCR were performed on infested field samples which showed a good agreement with our RT-iPCR results. Thus, RT-iPCR may be an invaluable tool for field diagnostics.en_US
dc.identifier.urihttps://hdl.handle.net/10133/5764
dc.language.isoen_USen_US
dc.proquest.subjectBiology [0306]en_US
dc.proquest.subjectMolecular biology [0307]en_US
dc.proquest.subjectBiochemistry [0487]en_US
dc.proquestyesYesen_US
dc.publisherLethbridge, Alta. : University of Lethbridge, Dept. of Biological Sciencesen_US
dc.publisher.departmentDepartment of Biological Sciencesen_US
dc.publisher.facultyArts and Scienceen_US
dc.relation.ispartofseriesThesis (University of Lethbridge. Faculty of Arts and Science)en_US
dc.subjectDissertations, Academicen_US
dc.subjectAphanomyces euteichesen_US
dc.subjectEnzyme-linked immunosorbent assayen_US
dc.subjectLegumes -- Diseases and pestsen_US
dc.subjectRoot rotsen_US
dc.titleDevelopment of a real time immuno-PCR assay for the detection of pea root rot causal agent, Aphanomyces euteichesen_US
dc.typeThesisen_US
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