Rapid and visual detection of tomato spotted wilt virus using recombinase polymerase amplification combined with lateral flow strips

2021 ◽  
Vol 57 ◽  
pp. 101727
Author(s):  
Hyo-Jeong Lee ◽  
In-Sook Cho ◽  
Ho-Jong Ju ◽  
Rae-Dong Jeong
2021 ◽  
Author(s):  
Kaiqiang Hao ◽  
Ming Gu ◽  
Miaoren Yang ◽  
Xinran Gao ◽  
Zihao Xia ◽  
...  

Abstract Tomato spotted wilt virus (TSWV) is one of the most economically destructive and scientifically challenging plant viruses, which has seriously affected the production of commercial crops. At present, there is no effective strategy to control this virus. Therefore, there is an urgent need for a rapid and simple method to detect TSWV, which is of great significance to prevent its spread. In this study, an isolate of TSWV (TSWV-LNTL) infecting pepper from Liaoning Province of northeast China was obtained. A phylogenetic tree based on neighbor-joining using coat protein (CP) gene was established. A rapid method for detecting TSWV by recombinase polymerase amplification (RPA) was established. The phylogenetic tree based on the nucleotide sequences of coat protein (CP) genes of different TSWV isolates showed that the genetic relationship of TSWV-LNTL was most closely related to that of TSWV-LX-Lettuce-12 (Yunnan) and TSWV-TSHL (Shandong) isolates in China. It can be finished at 39 °C for 20 min and then purified by heating at 65 °C for 10 min. The RPA primers were highly specific and no cross-reactivity was detected with other selected viruses infecting pepper. The results of sensitivity test revealed that the detection limit of RPA is 1.0 × 103 copies/μL, which was tenfold lower than that of PCR method. In addition, the RPA method was successfully applied to detect TSWV in field samples. These results reported the occurrence of TSWV on crop in Liaoning Province of northeast China and demonstrated that the established RPA assay provided an effective molecular diagnostic tool for the accurate and rapid detection of TSWV to prevent its spread.


Pathogens ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 320
Author(s):  
Alexander Nilon ◽  
Karl Robinson ◽  
Hanu R. Pappu ◽  
Neena Mitter

Tomato spotted wilt virus (TSWV) is the type member of the genus Orthotospovirus in the family Tospoviridae and order Bunyavirales. TSWV, transmitted by several species of thrips, causes significant disease losses to agronomic and horticultural crops worldwide, impacting both the yield and quality of the produce. Management strategies include growing virus-resistant cultivars, cultural practices, and managing thrips vectors through pesticide application. However, numerous studies have reported that TSWV isolates can overcome host-plant resistance, while thrips are developing resistance to pesticides that were once effective. RNA interference (RNAi) offers a means of host defence by using double-stranded (ds) RNA to initiate gene silencing against invading viruses. However, adoption of this approach requires production and use of transgenic plants and thus limits the practical application of RNAi against TSWV and other viruses. To fully utilize the potential of RNAi for virus management at the field level, new and novel approaches are needed. In this review, we summarize RNAi and highlight the potential of topical or exogenous application of RNAi triggers for managing TSWV and thrips vectors.


Nature ◽  
1964 ◽  
Vol 203 (4945) ◽  
pp. 671-672 ◽  
Author(s):  
RUPERT J. BEST ◽  
GERARD F. KATEKAR

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