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Improved Diagnostic Tools to Reliably Detect Modern Wheat Mosaic Virus Isolates
Matthew West1, Olive LoGrasso1, Robyn Roberts1
1Department of Agricultural Biology, Colorado State University, Fort Collins, CO 80523, U.S.A.
Plant Disease
|January 15, 2026
Summary
New primers and a loop-mediated isothermal amplification (LAMP) assay improve detection of Triticum mosaic virus (TriMV) and Wheat streak mosaic virus (WSMV) in wheat. These tools reliably identify new virus isolates, crucial for plant disease control.
Area of Science:
- Plant Pathology
- Virology
- Molecular Diagnostics
Background:
- Accurate plant disease diagnostics are vital for crop protection, but genomic variation in virus isolates challenges existing detection methods.
- Triticum mosaic virus (TriMV) and Wheat streak mosaic virus (WSMV) cause significant economic damage to wheat crops in the US Great Plains.
- Traditional PCR primers for TriMV and WSMV showed unreliability with Colorado isolates, leading to false negatives in a substantial portion of tested samples.
Purpose of the Study:
- To develop novel, universally applicable diagnostic tools for detecting emerging isolates of TriMV and WSMV in wheat.
- To enhance the efficiency, sensitivity, and specificity of viral detection compared to existing methods.
- To create high-throughput and accessible bioassays for reliable plant virus identification.
Main Methods:
- Comparative genomic analysis to identify conserved regions for primer design.
- Development and validation of new universal primers for WSMV and TriMV using quantitative PCR (qPCR).
- Design of a multiplex PCR assay for simultaneous detection of both viruses and a colorimetric loop-mediated isothermal amplification (LAMP) assay for WSMV and/or TriMV detection.
Main Results:
- New universal primers demonstrated 16-18% greater efficiency and 16% higher sensitivity in qPCR compared to established primers.
- The developed primers exhibited high specificity for WSMV and TriMV (E-values near zero) and low cross-reactivity with the wheat genome (E-values = 1.9-7.5).
- Both the multiplex PCR and LAMP assays proved reliable, specific, and highly sensitive in detecting WSMV and TriMV, including new Colorado isolates.
Conclusions:
- Modified primers and novel bioassays (multiplex PCR and LAMP) offer improved accuracy and reliability for detecting WSMV and TriMV.
- These enhanced diagnostic tools are essential for effective management of economically significant wheat viruses, especially in the face of evolving viral populations.
- The developed LAMP assay provides a user-friendly and sensitive method for rapid plant virus detection in diverse settings.
