Rapid determination of Phytophthora infestans sporangia using a surface plasmon resonance immunosensor.
Phytophthora infestans is the cause of late blight disease in potato and is an economically important pathogen worldwide. Early disease detection is important to implement disease control measures. In this study a surface plasmon resonance (SPR) immunosensor for detection of P. infestans sporangia is presented. The specificity of an existing mouse monoclonal antibody (phyt/G1470 mAb) against P. infestans was investigated in plate-trapped antigen ELISA and in subtractive inhibition ELISA. No or only limited cross-reactivity was observed against representatives having air-borne spores from Ascomycetes, Deuteromycetes as well as Basidiomycetes. phyt/G1470 mAb was incorporated in a subtractive inhibition SPR assay, consisting of a pre-incubation of mAb and sporangia, a centrifugation step to remove sporangia-bound phyt/G1470 mAb and quantification of remaining phyt/G1470 mAb by SPR. Good intra- and interday assay variability was observed and the assay had a detection limit of 2.2x10(6) sporangia/ml. Analysis time was 75 min, which is superior to existing P. infestans detection methods.
Skottrup
Peter
P
Danish Institute of Agricultural Sciences, Department of Integrated Pest Management, Research Centre Flakkebjerg, DK-4200 Slagelse, Denmark. peter.skottrup@gmail.com
Nicolaisen
Mogens
M
Justesen
Annemarie F
AF
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D051379:Mice; D008856:Microscopy, Fluorescence; D010838:Phytophthora / Q000276:immunology / Q000302:isolation & purification* / Q000502:physiology; D010935:Plant Diseases / Q000382:microbiology; D013172:Spores, Fungal / Q000276:immunology / Q000302:isolation & purification*; D020349:Surface Plasmon Resonance / Q000295:instrumentation / Q000379:methods*; D013997:Time Factors
D000911:Antibodies, Monoclonal