Monoclonal antibodies for the detection of Puccinia striiformis urediniospores.
The fungal pathogen Pst causes yellow rust disease in wheat plants leading to crop losses. The organism spreads by releasing wind-dispersed urediniospores from infected plants. In this study a library of novel monoclonal antibodies (mAbs) was developed against Pst urediniospores. Nine mAb-producing cell lines were cloned and their cross-reactivities characterised against a panel of airborne fungal spores representing genera commonly found in the same environment as Pst. Two specific mAbs were used to develop a competitive ELISA (Pst mAb4) and a subtractive inhibition ELISA (Pst mAb8). Standard curves for both assays had good intra- and interday reproducibility. The subtractive inhibition ELISA had greater sensitivity with a detection limit of 1.5 x 10(5) spores ml(-1). Cross-reactivity studies of Pst mAb8 in the subtractive inhibition ELISA, showed reaction with other Puccinia spores only, suggesting that common epitopes exist within this genus. The biosensor-compatible Pst mAb8 assay principle developed in this study has the potential to be implemented in future 'label-free' in-the-field systems for Pst detection.
Skottrup
Peter
P
Danish Institute of Agricultural Sciences, Department of Integrated Pest Management, DK-4200 Slagelse, Denmark. psk@biocentrum.dtu.dk
Frøkiaer
Hanne
H
Hearty
Stephen
S
O'Kennedy
Richard
R
Hejgaard
Jørn
J
Nicolaisen
Mogens
M
Justesen
Annemarie F
AF
D000818:Animals; D000908:Antibodies, Fungal / Q000096:biosynthesis / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D000918:Antibody Specificity; D001487:Basidiomycota / Q000276:immunology*; D000939:Epitopes / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D010935:Plant Diseases / Q000382:microbiology; D013172:Spores, Fungal / Q000276:immunology; D014908:Triticum / Q000382:microbiology
D000908:Antibodies, Fungal; D000911:Antibodies, Monoclonal; D000939:Epitopes