Generation of Synthetic Peptide-Specific Antibody for the Development of A Southern Rice Black-Streaked Dwarf Virus Diagnostic Test

Do Thi Hanh,Nguyen Anh Minh,Nguyen Duy Phuong,Nguyen Van Cuu,Pham Xuan Hoi,Phung Thi Thanh Huong

doi:10.31817/vjas.2021.4.3.08

Abstract

Southern rice black-streaked dwarf virus (SRBSDV) causes severe epidemical disease on rice with the infected area up to millions of hectares in South China and North and Central of Vietnam. So far, there are no effective, cheap, quick, and practicable methods for diagnosing SRBSDV. The conventional RT-PCR technique is the most popular method for detecting SRBSDV with high accuracy. However, it is hard to apply this method for large-scale SDBSDV diagnosis because of the requirements of expensive reagents and instruments, as well as complex procedures. Meanwhile, SRBSDV diagnostic techniques based on antigen detection have outstanding advantages due to their low cost, easy manipulation, and wide application possibility. Today, there are still no commercially available specific antibodies to SRBSDV. In a previous study, to develop the SRBSDV diagnostic technique by the ELISA technique, a SRBSDV specific antibody was generated by a recombinant P10 envelope protein (66kDa), which has a titer of 1:5,000. In this study, we continued to study the production of SRBSDV specific polyclonal antibodies from small antigen–rich peptides from the SRBSDV P10 envelope protein. The resulting purified antibody can specifically bind to the P10 protein and at the diluted concentration of 1:100,000 it can detect SRBSDV in infected rice samples via the dot-blot technique. Our research results open up new opportunities for proactive antibodies to develop a SRBSDV membrane rapid diagnostic kit.

Highlights

Southern rice black-streaked dwarf virus (SRBSDV), a member of the genus Fijivirus in the family Reoviridae, is one of the mostDo Thi Hanh et al (2021)dangerous harmful rice viruses and is distributed mainly in southern Asia including China, Japan, and Vietnam (Guo et al, 2008; Zhang et al, 2008; Ngo Vinh Vien et al, 2009; Cuong et al, 2010; Wang et al, 2010; Matshukura et al, 2013)
The P10 protein of SRBSDV is a common target in diagnostic assays of SRBSDV using specific antibodies (Zhang et al, 2013; Do Thi Hanh et al, 2015)
To create antibodies binding to the P10 protein of Vietnamese SRBSDV isolates, the peptide fragment antigen was screened according to whether it was located on the (i) antigenic or (ii) conservative region of the P10 protein

Summary

Introduction

Southern rice black-streaked dwarf virus (SRBSDV), a member of the genus Fijivirus in the family Reoviridae, is one of the mostDo Thi Hanh et al (2021)dangerous harmful rice viruses and is distributed mainly in southern Asia including China, Japan, and Vietnam (Guo et al, 2008; Zhang et al, 2008; Ngo Vinh Vien et al, 2009; Cuong et al, 2010; Wang et al, 2010; Matshukura et al, 2013). Southern rice black-streaked dwarf virus (SRBSDV), a member of the genus Fijivirus in the family Reoviridae, is one of the most. The segment S10 has a length of 1,797bp, containing 1,674 nucleotides and an ORF encoding proteins that form the outer shell of the virus molecule with a corresponding molecular weight of 62.6kDa. The P10 protein is highly conservative, and has been identified as having specific interactions with the vectors of SRBSDV; it plays an important role in the evolution of the virus. The P10 protein is highly conservative, and has been identified as having specific interactions with the vectors of SRBSDV; it plays an important role in the evolution of the virus For this reason, the P10 protein is often chosen as the target for developing SRBSDV diagnostic methods (Yin et al, 2011)

Methods

Results

Conclusion