Monoclonal Antibody-Based Serological Detection of Rice Stripe Mosaic Virus Infection in Rice Plants or Leafhoppers

Abstract

Rice stripe mosaic virus (RSMV) is a rhabdovirus recently found in southern part of China and can cause severe reduction in rice production. To establish serological methods for RSMV epidemiological studies and to establish a control strategy for this virus, we first purified RSMV virions from infected rice plants and then used them as an immunogen to produce four RSMV-specific monoclonal antibodies (MAbs) (i.e.,1D4, 4A8, 8E4 and 11F11). With these MAbs, we have developed a highly specific and sensitive antigen-coated plate enzyme-linked immunosorbent assay (ACP-ELISA), a Dot-ELISA and a Tissue print-ELISA for rapid detections of RSMV infection in rice plants or in leafhoppers. Our results showed that RSMV can be readily detected in RSMV-infected rice plant tissue crude extracts diluted at 1:20,971,520 (w/v, g/mL) through ACP-ELISA or diluted at 1:327,680 (w/v, g/mL) through Dot-ELISA. Both ACP-ELISA and Dot-ELISA can also be used to detect RSMV infection in individual RSMV viruliferous leafhopper (Recilia dorsalis) homogenate diluted at 1:307,200 and 1:163,840 (individual leafhopper/µL), respectively. Detection of RSMV infection in field-collected rice samples or in RSMV viruliferous leafhoppers indicated that the three serological methods can produce same results with that produced by RT-PCR (19 of the 33 rice samples and 5 of the 16 leafhoppers were RSMV-positive). We consider that the four MAbs produced in this study are very specific and sensitive, and the three new serological methods are very useful for detections of RSMV infection in rice plants or in leafhoppers and the establishment of the disease control strategies.

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Acknowledgements

Project was supported by the Ministry of Agriculture of China (No. 2016ZX08009003-001), the National Key Research and Development Program of China (No. 2016YFD0300706), the National Natural Science Foundation of China (No. 31571976), and the Earmarked Fund for China Agriculture Research System (No. nycytx-001).

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Contributions

LG and JW prepared the MAbs and developed the serological assays. RC performed RT-PCR and field sample detections. JH participated in the study design and wrote part of the manuscript. XZ and JW conceived the study and finalized the manuscript. All authors have read and approved the final manuscript.

Corresponding authors

Correspondence to Xueping Zhou or Jianxiang Wu.

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All authors declare that they have no conflict of interest.

Animal and Human Rights Statement

The animal experiments were performed in accordance with the Principles of the Helsinki accords and approved by the Animal Experimentation Ethics Committee of Zhejiang University, Hangzhou, China.

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Guo, L., Wu, J., Chen, R. et al. Monoclonal Antibody-Based Serological Detection of Rice Stripe Mosaic Virus Infection in Rice Plants or Leafhoppers. Virol. Sin. 35, 227–234 (2020). https://doi.org/10.1007/s12250-019-00186-1

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Keywords

  • Rice stripe mosaic virus (RSMV)
  • Serological test
  • Antigen-coated plate enzyme-linked immunosorbent assay (ACP-ELISA)
  • Dot-ELISA
  • Tissue print-ELISA