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

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Kaphle, Salina
University of Lethbridge. Faculty of Arts and Science
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Lethbridge, Alta. : University of Lethbridge, Dept. of Biological Sciences
Effective 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.
Dissertations, Academic , Aphanomyces euteiches , Enzyme-linked immunosorbent assay , Legumes -- Diseases and pests , Root rots