Development of a Dot-Elisa Assay for Diagnosis of Southern Rice Black-Streaked Dwarf Disease in the Field

Date Received: Aug 09, 2021

Date Published: Sep 30, 2022

DOI: https://doi.org/10.31817/vjas.2020.5.3.05

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Issue: Vol. 5 No. 3 (2022): Vietnam Journal of Agricultural Sciences

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ENGINEERING AND TECHNOLOGY

How to Cite:

Duy, L., Hanh, D., Huong, B., Hang, P., Cuong, H., Hoi, P., … Phuong, N. (2022). Development of a Dot-Elisa Assay for Diagnosis of Southern Rice Black-Streaked Dwarf Disease in the Field. Vietnam Journal of Agricultural Sciences, 5(3), 1563–1570. https://doi.org/10.31817/vjas.2020.5.3.05

Development of a Dot-Elisa Assay for Diagnosis of Southern Rice Black-Streaked Dwarf Disease in the Field

La Duc Duy (*) 1 , Do Thi Hanh 2 , Bui Thi Thu Huong 1 , Pham Thu Hang 3 , Ha Viet Cuong 1 , Pham Xuan Hoi 3 , Dam Quang Hieu 3   , Nguyen Duy Phuong 3

  • Corresponding author: phuongnd.bio@gmail.com
  • 1 Faculty of Biotechnology, Vietnam National University of Agriculture, Hanoi 131000, Vietnam
  • 2 Faculty of Chemical Technology, Hanoi University of Industry, Hanoi 143510, Vietnam
  • 3 Department of Molecular Pathology, Agricultural Genetics Institute, Hanoi 143510, Vietnam

    • Keywords

    Antibody, diagnosis, dot-ELISA, P10 protein, SRBSDV

    Abstract


    Outbreaks of the Southern rice black-streaked dwarf virus (SRBSDV) have caused significant losses in many rice-growing areas in Vietnam, especially in both North and Central Vietnam in recent years. To detect the virus, traditional reverse transcription polymerase chain reaction (RT-PCR) methodology and immunoassays are currently employed. RT-PCR is accurate but requires expensive chemicals and instruments, as well as complex procedures that limit its applicability for field tests. To develop a cheaper, simpler, and reliable SRBSDV diagnosis assay based on the dot-enzyme-linked immunosorbent assay (dot-ELISA) method, anti-SRBSDV polyclonal antibodies were produced by using the antigens derived from the P10 coat protein of SRBSDV, which was achieved from a previous study. The IgG antibody purified from the antiserum of recombinant P10-immunized mice by protein A-agarose affinity chromatography could specifically detect both the target protein and SRBSDV at a dilution of 1:100000. In the trial test of SRBSDV diagnosis, the dot-ELISA assay using the obtained anti-SRBSDV antibody showed an accuracy rate of 90.9% in comparison with the standard RT-PCR assay. These results are important premises for the large-scale application of dot-ELISA assay for SRBSDV diagnosis in order to protect rice crops against viral disease damage.

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