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Development and use of three monoclonal antibodies for the detection of rice black-streaked dwarf virus in field plants and planthopper vectors

文献类型： 外文期刊

作者： Wu, Jianxiang 1 ; Ni, Yuequn 1 ; Liu, Huan 1 ; Rao, Lixia 1 ; Zhou, Yijun 2 ; Zhou, Xueping 1 ;

作者机构： 1.Zhejiang Univ, Inst Biotechnol, State Key Lab Rice Biol, Hangzhou 310058, Zhejiang, Peoples R China

2.Jiangsu Acad Agr Sci, Inst Plant Protect, Nanjing 210014, Peoples R China

关键词： Rice black-streaked dwarf virus;Monoclonal antibody;Small brown planthopper;Antigen-coated-plate enzyme-linked immunosorbent assay;Dot enzyme-linked immunosorbent assay

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： 2013 年 10 卷

页码：

收录情况： SCI

摘要： Background: Rice black-streaked dwarf virus (RBSDV) causes great losses in rice, maize and wheat production in Asian countries. The use of serological methods for RBSDV detection depends on the availability of antibodies. In this study, three highly sensitive and specific murine monoclonal antibodies (MAbs) against RBSDV antigens were produced using crude extracts from tumors of RBSDV-infected maize as the immunogen, and two serological assays, antigen-coated-plate enzyme-linked immunosorbent assay (ACP-ELISA) and dot enzyme-linked immunosorbent assay (dot-ELISA) were developed for RBSDV detection. Results: All three MAbs reacted strongly and specifically with the crude extracts from RBSDV-infected plant and planthopper tissues. The detection endpoints of three MAbs (12E10, 18F10 and 5G5) in ACP-ELISA were respectively 1:40,960, 1:40,960, 1:81,920 (w/v, g mL(-1)) with the crude extract of infected maize, 1: 10,240, 1: 20,480, 1: 20,480 (w/v, g mL(-1)) with the crude extract of infected rice, 1: 5,120, 1: 10,240, 1: 10,240 (w/v, g mL(-1)) with the crude extract of infected wheat, 1: 9,600, 1: 9,600, 19,200 (individual planthopper/mu L) with the crude extract of infected planthopper. The newly developed ACP-ELISA could detect the virus in the infected maize, wheat, rice tissue crude extracts diluted at 1: 81,920, 1: 20,480, 1: 10,240 (w/v, g mL(-1)), respectively, and in individual viruliferous planthopper extract diluted at 1: 19200 (individual planthopper/mu L). The dot-ELISA was proved to detect the virus in the infected maize, wheat and rice tissue crude extracts diluted at 1: 320 (w/v, g mL(-1)), and in individual viruliferous planthopper extract diluted at 1: 1,600 (individual planthopper/mu L), respectively. Field plants (915) and planthopper samples (594) from five provinces of China were screened for the presence of RBSDV using the two developed serological assays. The results indicated that 338 of the 915 plant samples and 19 of the 594 planthopper samples were infected by RBSDV. Conclusions: The newly developed ACP-ELISA and dot-ELISA were highly sensitive and specific to detect RBSDV in field plant and planthopper samples. The field survey demonstrated that RBSDV is widespread in rice, maize and wheat crops in Jiangsu, Zhejiang, Shandong provinces of China.

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