Journal
FOOD AND AGRICULTURAL IMMUNOLOGY
Volume 34, Issue 1, Pages 10-20Publisher
TAYLOR & FRANCIS LTD
DOI: 10.1080/09540105.2023.2185568
Keywords
Tobacco mosaic virus; time-resolved fluorescence immunoassay; rapid diagnostic test; lateral flow immunoassay; field detection
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Tobacco mosaic virus (TMV) was detected in tobacco using a time-resolved fluorescence immunoassay (TRFIA) with a detection limit of 0.24 ng/mL. Monoclonal antibodies developed from TMV virions were conjugated to Eu3+ nanospheres. The specificity of the TRFIA platform was confirmed using ten species of tobacco-infecting phytopathogens. The TRFIA platform was successfully applied for on-site TMV testing in 10,000 tobacco leaf samples collected from five fields in Kunming, China.
Tobacco mosaic virus (TMV) causes a significant yield loss in solanaceous crops. Accurate and rapid diagnosis of TMV is an essential pre-requisite for proactive and tailored disease management strategies, filling the gap between more rigorous laws regarding pesticide use and maintaining crop production. Herein, we developed a time-resolved fluorescence immunoassay (TRFIA) to quantitatively detect TMV in tobacco. TMV virions were used as immunogens to develop monoclonal antibodies, which were then conjugated to Eu3+ nanospheres. The detection limit of the TRFIA platform was 0.24 ng/mL, with recovery rates in the range of 90.71-108.11% in spiked tobacco. The specificity was confirmed using ten species of tobacco-infecting phytopathogens. In-field applicability was implemented by screening for TMV in 10,000 tobacco leaf samples collected from five fields in Kunming, China. Overall, the proposed TRFIA platform is rapid, accurate, and end-user friendly, demonstrating its application in TMV onsite testing in tobacco.
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