Development of high-throughput quantum dot biosensor against Polymyxa species.
The plasmodiophoromycete Polymyxa betae and P. graminis are eukaryotic biotrophic parasites residing in the roots of chenopodiacae and gramineae plants. They are natural transmitting agents of several important plant viruses such as are beet necrotic yellow vein virus (BNYW), beet soil borne mosaic virus (BSBMV), wheat soil-borne mosaic virus (WSBMV). Developing advanced high-throughput diagnostic methods capable of accurate detection of these pathogens could assist with the screening programs and consequently with the development of disease-resistant germplasms. In the present study, a previously developed quantum dots (QDs) FRET-based nano-biosensor was upgraded to a high-throughput version. QDs were functionalized with a specific antibody against the P. betae's specific glutathione-S-transferase (GST) protein. On the other hand, GST was conjugated to Rhodamine dye. Ninety six-well polystyrene plates were used as the detection platform. The mutual affinity of the antigen and the antibody brought the CdTe QDs and rhodamine together close enough to allow the resonance dipole-dipole coupling required for fluorescence resonance energy transfer (FRET) to occur. The immunosensor constructed showed a high sensitivity and specificity of 100%, and was successfully used for high-throughput screening of 96 real samples with consistent results within the course of less than 30 minutes.