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Background <p>Sugarcane yellow leaf virus (SCYLV) is an important pathogen that induces severe leaf yellowing and substantial yield loss in sugarcane cultivation. The absence of resistant cultivars underscores the urgent need for large-scale detection techniques to control SCYLV spread.</p> Methods <p>We employed single B-cell technology to generate a specific monoclonal antibody (mAb) against the SCYLV coat protein (CP). Using this mAb, we established a tissue blot immunoassay (TBIA) for viral detection based on imprinting sugarcane leaf midrib tissues onto nitrocellulose membranes. The complete light and heavy chain sequences of the antibody have been determined.</p> Results <p>The developed TBIA demonstrated superior efficiency compared to conventional RT-PCR, and is capable of processing 300 samples within 24&#xa0;h at a low cost of 0.28 USD per test. Validation with 40 field samples showed 100% concordance with RT-PCR results and only one discrepancy compared to RT-MIRA-CRISPR/Cas12a assays.</p> Conclusion <p>In this study, we report the first application of single B-cell technology for SCYLV diagnostics, provides a rapid, reliable, and cost-effective TBIA method for SCYLV detection, which facilitates early warning and enables integrated prevention and control of sugarcane yellow leaf disease.</p> Graphical Abstract <p></p>

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Development and application of monoclonal antibody-based tissue-blot immunoassay for rapid, specific, and sensitive detection of sugarcane yellow leaf virus

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年1月10日

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  • Ting Wang,
  • Anzhen Li,
  • Shuai Gao,
  • Jie Guo,
  • Dantong Yang,
  • Liping Xu,
  • Jinlong Guo

摘要

Background

Sugarcane yellow leaf virus (SCYLV) is an important pathogen that induces severe leaf yellowing and substantial yield loss in sugarcane cultivation. The absence of resistant cultivars underscores the urgent need for large-scale detection techniques to control SCYLV spread.

Methods

We employed single B-cell technology to generate a specific monoclonal antibody (mAb) against the SCYLV coat protein (CP). Using this mAb, we established a tissue blot immunoassay (TBIA) for viral detection based on imprinting sugarcane leaf midrib tissues onto nitrocellulose membranes. The complete light and heavy chain sequences of the antibody have been determined.

Results

The developed TBIA demonstrated superior efficiency compared to conventional RT-PCR, and is capable of processing 300 samples within 24 h at a low cost of 0.28 USD per test. Validation with 40 field samples showed 100% concordance with RT-PCR results and only one discrepancy compared to RT-MIRA-CRISPR/Cas12a assays.

Conclusion

In this study, we report the first application of single B-cell technology for SCYLV diagnostics, provides a rapid, reliable, and cost-effective TBIA method for SCYLV detection, which facilitates early warning and enables integrated prevention and control of sugarcane yellow leaf disease.

Graphical Abstract