Development of a sensitive Luminex xMAP-based microsphere immunoassay for specific detection of Iris yellow spot virus
文献类型: 外文期刊
第一作者: Yu, Cui
作者: Yu, Cui;Yang, Cuiyun;Song, Shaoyi;Yu, Zixiang;Zhou, Xueping;Wu, Jianxiang;Zhou, Xueping
作者机构:
关键词: Iris yellow spot virus;Antibody;Microsphere immunoassay;Detection
期刊名称:VIROLOGY JOURNAL ( 影响因子:4.099; 五年影响因子:3.719 )
ISSN: 1743-422X
年卷期: 2018 年 15 卷
页码:
收录情况: SCI
摘要: Background: Iris yellow spot virus (IYSV) is an Orthotospovirus that infects most Allium species. Very few approaches for specific detection of IYSV from infected plants are available to date. We report the development of a high-sensitive Luminex xMAP-based microsphere immunoassay (MIA) for specific detection of IYSV. Results: The nucleocapsid (N) gene of IYSV was cloned and expressed in Escherichia coli to produce the His-tagged recombinant N protein. A panel of monoclonal antibodies (MAbs) against IYSV was generated by immunizing the mice with recombinant N protein. Five specific MAbs (16D9, 11C6, 7F4, 12C10, and 14H12) were identified and used for developing the Luminex xMAP-based MIA systems along with a polyclonal antibody against IYSV. Comparative analyses of their sensitivity and specificity in detecting IYSV from infected tobacco leaves identified 7F4 as the best-performed MAb in MIA. We then optimized the working conditions of Luminex xMAP-based MIA in specific detection of IYSV from infected tobacco leaves by using appropriate blocking buffer and proper concentration of biotin-labeled antibodies as well as the suitable ratio between the antibodies and the streptavidin R-phycoerythrin (SA-RPE). Under the optimized conditions the Luminex xMAP-based MIA was able to specifically detect IYSV with much higher sensitivity than conventional enzyme-linked immunosorbent assay (ELISA). Importantly, the Luminex xMAP-based MIA is time-saving and the whole procedure could be completed within 2.5 h. Conclusions: We generated five specific MAbs against IYSV and developed the Luminex xMAP-based MIA method for specific detection of IYSV in plants. This assay provides a sensitive, high-specific, easy to perform and likely cost-effective approach for IYSV detection from infected plants, implicating potential broad usefulness of MIA in plant virus diagnosis.
分类号: Virology
- 相关文献
作者其他论文 更多>>
-
RNA modifications in plant biotic interactions
作者:Ge, Linhao;Pan, Fuan;Jia, Mingxuan;He, Hao;Shan, Hongying;Zhou, Xueping;Li, Fangfang;Pan, Fuan;Zhou, Xueping;Pott, Delphine M.;Wang, Aiming
关键词:RNA modifications; N6-methyladenosine; m6A; m6A core components; plant biotic stress; plant- microbe interactions; plant-insect interactions
-
Soybean stay-green associated geminivirus: A serious threat to soybean production in China
作者:He, Hao;Li, Hao;Zhou, Xueping;Li, Fangfang;He, Hao;Cui, Xiaoyan;Wang, Yaqin;Zhou, Xueping;Xu, Yi
关键词:Soybean; Soybean stay-green associated geminivirus; (SoSGV); Soybean stay-green syndrome; Viral transmission; Viral epidemiology
-
Multiplex lateral flow immunoassay for simultaneous detection and specific discrimination of Xoo and Xoc in rice plants
作者:Zhang, Cui;Zhang, Xi;Liu, Nairu;Dong, Jie;Mao, Weijia;Zhou, Xueping;Wu, Jianxiang;Zhang, Cui;Wu, Jianxiang;Liu, Zhaoli;Zhou, Xueping
关键词:Multiplex lateral flow immunoassay; Simultaneous detection; AuNPs; Xoo; Xoc
-
Editing of an antiviral host factor boosts plant growth and yield of plant viral vector-mediated heterologous protein expression
作者:Fang, Zhu;Zhao, Xinru;Du, Min;Xu, Xinyi;Zhou, Hui;Zhou, Xueping;Yang, Xiuling;Guo, Wei;Zhou, Xueping
关键词:Nicotiana benthamiana; antiviral host factor; genome editing; plant biofactory
-
Developing glycosylase-based T-to-G and C-to-K base editors in rice
作者:Kuang, Yongjie;Wu, Xuemei;Liu, Meijie;Yan, Fang;Zhou, Xueping;Zhou, Huanbin;Ren, Bin;Liu, Meijie;Ma, Dongfang;Yan, Fang;Zhou, Huanbin;Zhou, Xueping
关键词:CRISPR/Cas9; thymine DNA glycosylase; cytosine DNA glycosylase; base editing; rice
-
Cucumber green mottle mosaic virus encodes additional small proteins with specific subcellular localizations and virulence function
作者:Gong, Pan;Gao, Mengxin;Chen, Yalin;Zhang, Mingzhen;Hu, Xiaohua;Zhao, Siwen;Pan, Mengjiao;Cao, Buwei;Zhou, Xueping;Li, Fangfang;Gao, Mengxin;Zhou, Xueping;Huang, Yucong;Shen, Qingtang;Zhang, Hui;Liu, Yong;Liu, Yong;Lozano-Duran, Rosa;Wang, Aiming
关键词:positive-sense single-stranded RNA viruses; tobamoviruses; cucumber green mottle mosaic virus; small ORFs; peroxisomes; virus pathogenicity
-
Geminivirus βV1 protein activates bZIP17/28mediated UPR signaling to facilitate viral pathogenicity but its activity is attenuated by autophagic degradation in plants
作者:Hu, Tao;Li, Chenyang;Liu, Hui;Su, Chenlu;Wang, Yaqin;Zhou, Xueping;Hu, Tao;Li, Fangfang;Zhou, Xueping;Li, Chenyang
关键词:unfolded protein response; autophagy; geminivirus; beta V1; ER stress; ATG18a