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Updated: Jan 29, 2026

High-Throughput Automated Multiplex Immunofluorescence Assays for Translational Research
Published on: June 10, 2025
An Immunofluorescence Assay to Detect Urediniospores of Phakopsora pachyrhizi
Fulya Baysal-Gurel1, Melanie L Lewis Ivey1, Anne Dorrance1
1Department of Plant Pathology, The Ohio State University, Ohio Agricultural Research and Development Center, Wooster.
Abstract:
An indirect immunofluorescence spore assay (IFSA) was developed to detect urediniospores of Phakopsora pachyrhizi, utilizing rabbit polyclonal antisera produced in response to intact nongerminated (SBR1A) or germinated (SBR2) urediniospores of P. pachyrhizi. Both antisera were specific to Phakopsora spp. and did not react with other common soybean pathogens or healthy soybean leaf tissue in enzyme-linked immunosorbent assay (ELISA). SBR1A and SBR2 bound to P. pachyrhizi and P. meibomiae urediniospores were detected with goat anti-rabbit Alexa Fluor 488-tagged antiserum using a Leica DM IRB epifluorescent microscope with an I3 blue filter (excitation 450 to 490 nm, emission 515 nm). The assay was performed on standard glass microscope slides; double-sided tape was superior to a thin coating of petroleum jelly both in retaining spores and in immunofluorescence. The IFSA was used to confirm the identity of P. pachyrhizi urediniospores captured on glass slides from passive air samplers from Georgia, Kentucky, and Ohio during 2006.
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