快速灵敏的甘蔗线条花叶病毒免疫学检测体系的建立

Rapid and Sensitive Identification of the SCSMV-infected Sugarcane Based on Immuno-detections

甘蔗花叶病广泛存在于我国甘蔗种植区,严重影响甘蔗产业的高质量发展。近年来甘蔗线条花叶病毒在蔗区肆虐,尽管针对其的血清学检测技术已经建立,但是快速、准确、高通量的检测方法亟待发掘。本研究制备了S CSMV CP的抗血清,特异性高,与引起甘蔗花叶病的另两种病原(高粱花叶病毒和甘蔗花叶病毒)间没有血清学交叉反应。基于该多克隆抗体,建立了直接抗原包被的ELISA、斑点杂交、Western blot和基于多抗的免疫试纸条检测技术。开发的免疫试纸条检测技术能快速、准确、高通量应用于田间病毒鉴定。本文提供了基于血清学的快速、准确、高通量,且便捷的甘蔗线条花叶病毒检测技术,有助于我国蔗区甘蔗花叶病的监测与防控。

Sugarcane mosaic disease wildly exists in sugarcane-cultivating fields and causes huge impact on sugar industry in China.Sugarcane streak mosaic virus(SCSMV),has been noted to become widely spread in recent years.Although serology-based detection techniques for SCSMV are established,rapid,accurate,and high-throughput detection methods are much less developed.Here,the PAb-SCSMVCP(Polyclonal antibody of SCSMVCP)was produced and its specificity was tested.There is no serological cross-reaction observed with other viruses that cause sugarcane mosaic disease too.Based on the PAb-SCSMVCP,the immune-detection methods,including double antibody sandwich(DAS)ELISA,dot blotting,and western blotting,were successfully deve-loped.Besides,the colloidal gold immunochromatographic strips were well-prepared and successfully used for rapid and accurate SCSMV detection in the wild field for the first time.Our studies provide serology based high-throughput,rapid,sensitive and quick diagnose methods for SCSMV detection in Chinese sugarcane-cultivating regions at first,which might contribute to the sugarcane mosaic disease control in China.