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Methodology for antibody preparation and detection of southern rice black-streaked dwarf virus

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Abstract

Southern rice black-streaked dwarf virus (SRBSDV) is a member of the genus Fijivirus of the family Reoviridae that causes a devastating disease in rice. Seven polypeptide fragments of the Putative uncharacterized protein (Pup) and the P10 proteins of SRBSDV were designed, synthesized, and used to immunize rabbits. Titers of polyclonal antibodies against the seven peptides were examined using indirect enzyme-linked immunosorbent assay (ELISA), and their specificities were investigated using western blotting. Indirect dot-immunobinding assay (DIBA) was also carried out at different dilutions against an antigen (rice extract). Antibody-1, which had the highest selectivity and titer, was then used to examine rice samples suspected of being infected with SRBSDV that were collected for over two years in different areas of China, using DIBA. Our results indicate that antibody-1 has the advantages of reliability, high sensitivity, and high specificity. Use of this antibody can help facilitate identification of the virus and its distribution in rice-growing areas where it causes significant problems.

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Acknowledgments

The authors gratefully acknowledge the financial support of the National Key Program for Basic Research (Nos. 2010CB 126105 and 2012CB722503), the Special Fund for Agro-scientific Research in the Public Interest (No. 201203022), and the National Natural Science Foundation of China (No. 21132003). We thank Prof. Qi Fang (Biotechnology and Genetic Germplasm Resources Institute, Yunnan Academy of Agricultural Sciences) and Prof. Aoshuang Chang (Guiyang Medical College), who carried out the ISEM experiments reported in this study.

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The authors declare no conflicts of interest related to this manuscript.

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Authors and Affiliations

  1. State Key Laboratory Breeding Base of Green Pesticide and Agricultural Bioengineering, Key Laboratory of Green Pesticide and Agricultural Bioengineering, Guizhou University, Ministry of Education, Guiyang, 550025, People’s Republic of China

    Zhuo Chen, Chengjun Yin, Jiaju Liu, Zhenchao Wang, Dandan Yu, Liang Bi, Linhong Jin, Song Yang & Baoan Song

  2. School of Chemistry and Chemical Engineering, South China University of Technology, Guangzhou, 510640, People’s Republic of China

    Mengjiao Zeng

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  1. Zhuo Chen
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  2. Chengjun Yin
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  3. Jiaju Liu
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Correspondence to Baoan Song.

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Chen, Z., Yin, C., Liu, J. et al. Methodology for antibody preparation and detection of southern rice black-streaked dwarf virus. Arch Virol 157, 2327–2333 (2012). https://doi.org/10.1007/s00705-012-1430-8

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  • DOI: https://doi.org/10.1007/s00705-012-1430-8

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