A rapid detection of tomato yellow leaf curl virus using recombinase polymerase amplification‐lateral flow dipstick assay

• Published in: Letters in applied microbiology Vol. 74; no. 5; pp. 640 - 646
• Main Authors: Zhou, Y., Zheng, H.Y., Jiang, D.M., Liu, M., Zhang, W., Yan, J.Y.
• Format: Journal Article
• Language: English
• Published: England Oxford University Press 01.05.2022
• Subjects: Amplification; Deoxyribonucleic acid; DNA; field detection; Instrumentation; lateral flow dipstick; Leaf-curl; Leaves; Plant diseases; Recombinase; recombinase polymerase amplification; tomato yellow leaf curl virus; Tomatoes; Viruses; Yellow leaf; lateral flow dipstick; recombinase polymerase amplification; field detection; tomato yellow leaf curl virus
• ISSN: 0266-8254; 1472-765X
• DOI: 10.1111/lam.13611

Tomato yellow leaf curl disease which is caused by Tomato yellow leaf curl virus (TYLCV) is economically important and a widely spread tomato disease in China. Rapid and accurate detection methods are important in the control TYLCV. Here, a rapid method was developed to identify TYLCV on the basis of recombinase polymerase amplification (RPA) that can be visualized in 5 min using lateral flow dipsticks. The sensitivity and the specificity of this method were evaluated. This method can detect 0·5 pg DNA after 30 min at 37°C without any expensive instrumentation. In addition, it showed higher sensitivity than a PCR method when purified DNA was used. Moreover, the TYLCV was specifically detected, whereas other viruses infecting tomato produced negative results. The crude tomato extracts used in this assay has potential application in minimally equipped plant clinic laboratories. This method will facilitate the early and rapid detection of TYLCV for the timely application of control measures. Significance and Impact of the Study: Tomato yellow leaf curl virus (TYLCV) cause tomato yellow leaf curl disease, which can result in severe damage to tomato production worldwide. Plants infected with TYLCV during the early growth stages frequently suffer yield losses of up to 100%. Rapid and accurate detection method for TYLCV is important for effective control of the disease. In this study, a RPA‐LFD method for rapid detection of TYLCV was developed. Compared to the conventional PCR method, the RPA‐LFD method combined with simple equipment is rapid, accurate and practical for field testing and survey of the occurrence of TYLCV.