MARA National Center for Soybean Improvement, Nanjing Agricultural University, Weigang 1, Nanjing 210095, China.
College of Agronomy, Henan Agricultural University, Zhengzhou 450046, China.
Int J Mol Sci. 2022 Aug 21;23(16):9457. doi: 10.3390/ijms23169457.
Soybean is an important grain and oil crop worldwide; however, the yield and seed quality of which are seriously affected by (SMV). As efficient detection technology is crucial for the field management of SMV, novel immunological detection methods were developed in the present study. According to the phylogenetic analysis, the CP coding sequence of SMV-SC7 was selected for the prokaryotic expression of the recombinant SMV-CP. Purified SMV-CP was used for the development of polyclonal antibodies (PAb) against the SMV-CP (PAb-SMV-CP) and monoclonal antibodies (MAb) against SMV-CP (MAb-SMV-CP). Subsequently, the PAb-SMV-CP was used for the development of a novel DAS- quantitative ELISA (DAS-qELISA) kit, of which the sensitivity was greater than 1:4000, and this could be used for the quantitative detection of SMV in China. Meanwhile, the MAb-SMV-CP was labeled with colloidal gold, and then was used for the development of the SMV-specific gold immunochromatography strip (SMV-GICS). The SMV-GICS gives accurate detection results through observed control lines and test lines in 5 to 10 min, sharing the same sensitivity as RT-PCR, and can be used for rapid, accurate and high-throughput field SMV detection. The DAS-qELISA kit and the SMV-GICA strip developed in this study are SMV-specific, sensitive, cheap and easy to use. These products will be conducive to the timely, efficient SMV epidemiology and detection in major soybean-producing regions in China and abroad.
大豆是世界范围内重要的粮食和油料作物,但其产量和种子品质受到大豆花叶病毒 (SMV) 的严重影响。由于有效的检测技术对于 SMV 的田间管理至关重要,因此本研究开发了新型免疫检测方法。根据系统进化分析,选择 SMV-SC7 的 CP 编码序列进行 SMV-CP 的原核表达。纯化的 SMV-CP 用于制备针对 SMV-CP 的多克隆抗体 (PAb-SMV-CP) 和单克隆抗体 (MAb-SMV-CP)。随后,将 PAb-SMV-CP 用于开发新型 DAS-定量 ELISA (DAS-qELISA) 试剂盒,其灵敏度大于 1:4000,可用于中国 SMV 的定量检测。同时,将 MAb-SMV-CP 标记胶体金,然后用于开发 SMV 特异性金免疫层析试纸条 (SMV-GICS)。SMV-GICS 通过在 5 至 10 分钟内观察控制线和检测线,给出准确的检测结果,与 RT-PCR 具有相同的灵敏度,可用于快速、准确和高通量的田间 SMV 检测。本研究开发的 DAS-qELISA 试剂盒和 SMV-GICA 试纸条具有 SMV 特异性、灵敏度高、价格低廉且易于使用的特点。这些产品将有利于及时、高效地开展国内外主要大豆产区的 SMV 流行病学和检测工作。