Abstract
This paper first time describes Surface Plasmon Resonance (SPR) based immuno-sensing system for detection of fungal teliospores of Karnal Bunt (KB) disease of wheat incited by Tilletia indica. The results of present study were compared with the results of lab scale ELISA developed earlier in our lab. The approach involves the use of a rabbit polyclonal (anti-teliospore) antibody and a SPR sensor for label-free detection of teliospores from the model organism Tilletia indica. When the interaction between binder (anti-teliospore antibody) and teliosporic antigen(s) as analyte occurs at the sensor surface, a sensorgram was obtained in real time by plotting the signal against time. Under optimised assay conditions as few as 5 teliospores were detected by ELISA which justifies the greater affinity of raised anti-teliospore antibodies for its antigen. The sensitivity of each immunosensor constructed with the different amounts of antigen solution is determined by sensogram analysis showed detection sensitivity as low as 625 pg equivalent to 2.5 teliospores over the sensor surface by getting an angle of dip indicating greater affinity of raised anti-teliospore antibodies. Thus, SPR based affinity sensor showed higher sensitivity due to greater signal response. The results of cross reactivity studies using different related fungal spores/teliospores showed almost differential SPR response indicating cross reaction using immunosensing system. The cross-reactivity of anti-teliospore antibodies with related fungal spores/teliospores can be explained due to its polyclonal nature and also due to antigenic homology amongst pathogens used in present study.
Highlights
Wheat (Triticum aestivum) is one of the most important food crops of the world
The results showed that interaction of teliosporic antigen-antiteliosporic antibody is more effective at pH 7.5 and thereby resulted in more angle change, pH 7.5 was chosen for further studies
This paper describes an optimization of Surface Plasmon Resonance (SPR) based immunosensing system for Karnal Bunt (KB) diagnosis using anti-teliosporic antibody (1:500 dilution) immobilized on a Self-Assembled Monolayer (SAM)
Summary
Wheat (Triticum aestivum) is one of the most important food crops of the world. In India, the wheat is affected by a number of diseases. Smuts, powdery mildew, loose smut, leaf blight and Karnal bunt continue to be the limiting factor in increasing wheat yield Amongst these diseases, Karnal bunt is the major disease of wheat (Triticum aestivum) and caused by the sporadic fungus, Tilletia indica (Syn Neovosia indica). Karnal Bunt (KB), called partial bunt, was first detected at Karnal (Haryana) by Mitra [1], and the name It affects durum wheat and triticale which result in reduced crop yield and poor grain quality. It gives off a fishy odour due to the presence of trimethylamine secreted by teliospores. As at present none of the control measures had been proven to be satisfactory for the disease management, it becomes inevitable to develop suitable detection system for seed health testing of wheat lots for strengthening seed certification and plant quarantine measures [2]
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