Ultrasensitive Nano-gold Labelled, Duplex Lateral Flow Immunochromatographic Assay for Early Detection of Sugarcane Mosaic Viruses

Sugarcane is one of the important food and bioenergy crops, cultivated all over the world except European continent. Like many other crops, sugarcane production and quality are hampered by various plant pathogens, among them viruses that infect systemically and cause severe impact to cane growth. The viruses are efficiently managed by their elimination through tissue culture combined with molecular diagnostics, which could detect virus titre often low at 10 g mL. To harmonize the virus diagnostics by molecular methods, we established a nanocatalysis-based high sensitive lateral flow immunochromatographic assay (LFIA) simultaneously to detect two major sugarcane viruses associated with mosaic disease in sugarcane. LFIA is known for poor sensitivity and stability with its signalling conjugates. However, we synthesized positively charged Cysteamine-gold nanoparticles and used them to prepare highly stable to sensitive immunoconjugates and as a colourimetric detection label. Further nanogold signal enhancement was performed on LFIA to obtain a high detection sensitivity, which is higher than the conventional immunoassays. The linear detection range of the nano-LIFA was 10 to 10 g mL, and with the signal enhancement, the LOD reached up to 10 g ml. This research paper provides relative merits and advancement on nano-LFIA for specific detection of sugarcane viruses in sugarcane for the first time.