Scholarlink

<p>Tomato brown rugose fruit virus (ToBRFV) is a single-stranded positive-sense RNA virus that targets tomato and pepper plants and is causing significant damage to crops in some regions of the world. ToBRFV is a highly contagious virus that is stable and rapidly spreads by mechanical methods and seeds. As a result, it may spread both locally and over long distances, and it is now recognized as a pandemic in plants. This study investigates the effectiveness of the systems CRISPR-Cas12a and CRISPR-Cas9, in conjugation with recombinase polymerase amplification (RPA), to detect ToBRFV in tomato plant samples collected from the field. The <i>trans</i>-cleavage activity of both nucleases, Cas12a and Cas9, was exploited to process a probe labelled with fluorescein and biotin to be resolved on a lateral flow device, thereby enabling a visual readout. We were able to detect the RNA genome of the virus in about 1&#xa0;h at a low constant temperature. These results could pave way to offer a rapid, sensitive, and specific method for on-site detection of ToBRFV.</p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Detection of tomato brown rugose fruit virus through CRISPR-Cas12a and CRISPR-Cas9 systems

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2025年7月15日

查看全文
  • Masoud Besati,
  • Mohammad Reza Safarnejad,
  • Atousa Aliahmadi,
  • Mohsen Farzaneh,
  • Raúl Ruiz,
  • Roser Montagud-Martínez,
  • Guillermo Rodrigo,
  • Hasan Rafati

摘要

Tomato brown rugose fruit virus (ToBRFV) is a single-stranded positive-sense RNA virus that targets tomato and pepper plants and is causing significant damage to crops in some regions of the world. ToBRFV is a highly contagious virus that is stable and rapidly spreads by mechanical methods and seeds. As a result, it may spread both locally and over long distances, and it is now recognized as a pandemic in plants. This study investigates the effectiveness of the systems CRISPR-Cas12a and CRISPR-Cas9, in conjugation with recombinase polymerase amplification (RPA), to detect ToBRFV in tomato plant samples collected from the field. The trans-cleavage activity of both nucleases, Cas12a and Cas9, was exploited to process a probe labelled with fluorescein and biotin to be resolved on a lateral flow device, thereby enabling a visual readout. We were able to detect the RNA genome of the virus in about 1 h at a low constant temperature. These results could pave way to offer a rapid, sensitive, and specific method for on-site detection of ToBRFV.