Plant Pathology Section, Division of Crop Protection, ICAR-Sugarcane Breeding Institute, Coimbatore, 641 007, India.
ICAR-National Bureau of Plant Genetic Resources, Regional Station, Hyderabad, 500030, India.
Sci Rep. 2022 Mar 9;12(1):4144. doi: 10.1038/s41598-022-07950-6.
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.
甘蔗是一种重要的粮食和生物能源作物,除了欧洲大陆以外,在世界各地都有种植。与许多其他作物一样,甘蔗的生产和质量受到各种植物病原体的影响,其中病毒会系统感染并严重影响甘蔗的生长。通过组织培养结合分子诊断可以有效地控制这些病毒,这种方法可以检测到通常浓度很低的 10 g/mL 的病毒载量。为了通过分子方法协调病毒诊断,我们建立了一种基于纳米催化的高灵敏度横向流动免疫层析分析(LFIA),同时检测与甘蔗花叶病相关的两种主要甘蔗病毒。LFIA 的信号结合物通常存在灵敏度和稳定性差的问题。然而,我们合成了带正电荷的半胱胺金纳米颗粒,并将其用于制备高稳定性和高灵敏度的免疫结合物,以及作为比色检测标记。进一步在 LFIA 上进行纳米金信号增强,以获得高检测灵敏度,其灵敏度高于传统免疫分析。纳米 LFIA 的线性检测范围为 10 到 10 g/mL,通过信号增强,LOD 可达到 10 g/ml。本研究论文首次提供了用于特异性检测甘蔗病毒的纳米 LFIA 的相对优点和进展。