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 and Nguyen Duy Phuong 3

1Faculty of Biotechnology, Vietnam National University of Agriculture, Hanoi 131000, Vietnam
2Faculty of Chemical Technology, Hanoi University of Industry, Hanoi 143510, Vietnam
3Department of Molecular Pathology, Agricultural Genetics Institute, Hanoi 143510, Vietnam
Received: Aug 9, 2021 /
Revised: Sep 30, 2022 /
Published: Sep 30, 2022

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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.

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

Article Details

How to Cite
Duy, L., Hanh, D., Huong, B., Hang, P., Cuong, H., Hoi, P., Hieu, D., & 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

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