Establishment of a Serology- and Molecular-Combined Detection System for Youcai Mosaic Virus and Its Application in Various Host Plants
2024
Chenwei Feng | Yanhong Hua | Duxuan Liu | Haoyu Chen | Mingjie Wu | Jing Hua | Kun Zhang
The youcai mosaic virus (YoMV) can infect a diverse array of crop species, such as <i>Raphanus sativus</i>, <i>Brassica napus</i>, <i>Solanum nigrum</i>, and <i>Rehmannia glutinosa</i>, causing substantial economic damage. This study aimed to develop a rapid, sensitive, and economical diagnostic method for YoMV. We successfully expressed and purified the recombinant His-CP<sup>YoMV-YZ</sup> protein in <i>E. coli</i> BL21, which was used to immunize New Zealand White rabbits, generating high-titer polyclonal antibodies (PAb-CP<sup>YoMV-YZ</sup>). Additionally, a serological-based reverse transcription loop-mediated isothermal amplification (S-RT-LAMP) assay was refined, combining serological and molecular detection techniques to enhance practicality. Utilizing PAb-CP<sup>YoMV-YZ</sup>, we developed four techniques for detecting YoMV: Western blot, dot immunoblotting assay, enzyme-linked immunosorbent assay (ELISA), and S-RT-LAMP. YoMV isolates from various regions and hosts were analyzed. The results indicated that PAb-CP<sup>YoMV-YZ</sup> was highly effective in detecting YoMV across a range of hosts and isolates from diverse regions. This study fills an important gap in the serological detection of YoMV and provides a practical tool for on-site diagnosis and control of YoMV infection.
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