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Comparison between gold nanoparticles and FITC as the labelling in lateral flow immunoassays for rapid detection of Ralstonia solanacearum

Ma, Lan, Wang, Shujuan, Xu, Dongpo, Xie, Manman, Ding, Chengchao, Tian, Yachen, Qiu, Jingxuan, Wang, Xiang, Dong, Qingli, Liu, Qing
Food and agricultural immunology 2018 v.29 no.1 pp. 1074-1085
Casuarina, Ralstonia solanacearum, bacterial wilt, cell fusion, cross reaction, detection limit, fluorescein, ginger, immunization, immunoaffinity chromatography, immunoassays, isothiocyanates, monitoring, monoclonal antibodies, nanogold, point-of-care testing, rapid methods, screening, tomatoes
Early prevention and monitoring of Ralstonia solanacearum (Rs) on nightshade is extremely critical for potent prevention and treatment of bacterial wilt disease. We developed two kinds of lateral flow immunochromatographic strip (LFICS) using gold nanoparticles (GNPs) and fluorescein isothiocyanate (FITC) for rapid diagnosis of plant bacterial wilt, caused by Ralstonia solanacearum (Rs), which both were based on double antibody sandwich method. Three monoclonal antibodies (McAb) highly specific to Rs were obtained by cell fusion and screening after four times of immunization with the strain GIM 1.76. Under optimized conditions, the limit of detection (LOD) with GNPs and FITC were 10⁴ colony forming units per millilitre (CFU/mL) and 10⁶ CFU/mL respectively, then the range detection correspondingly were 10⁴–10⁹ CFU/mL and 10⁶–10⁹ CFU/mL. On top of that, the developed immunoassay could be used for spot and point-of-care testing Rs on tomato seed, ginger seed, casuarina seed and display no cross-reaction with 43 other phytopathogenic strains that also could infect plants so as to cause plants diseases. These results demonstrated that we sucessfully developed two simple, effective and sensitive strips for monitoring bacterial wilt infection. Meanwhile, this is the first report about LFICS labelled by FITC for rapid detection of Rs, and it is the first time conformed that the sensitive of the LFICS with GNPs is significantly higher than FITC.