Deraniyagala, A S and Roy, Avijit and Tallury, S and Sudini, H K and Culbreath, A K and Bag, S (2025) Development of a Multiplex TaqMan Assay for Rapid Detection of Groundnut Bud Necrosis Virus: A Quarantine Pathogen in the USA. Viruses, 17 (4). pp. 1-14. ISSN 1999-4915
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Abstract
Groundnut bud necrosis orthotospovirus (GBNV), a tripartite single-stranded RNA virus, poses a significant threat to United States agriculture. GBNV is a quarantine pathogen, and its introduction could lead to severe damage to economically important crops, such as groundnuts, tomatoes, potatoes, peas, and soybeans. For the rapid and accurate detection of GBNV at points of entry, TaqMan reverse transcriptase–quantitative polymerase chain reaction (RT-qPCR) assays were developed and the results validated using conventional reverse transcriptase–polymerase chain reaction (RT-PCR) followed by Sanger sequencing. These assays target highly conserved regions of the nucleocapsid (NP) and movement (MP) proteins within the viral genome. Multiplex GBNV detection assays targeting the NP and MP genes, as well as an internal control plant gene, ACT11, showed efficiency rates between 90% and 100% and R2 values of 0.98 to 0.99, indicating high accuracy and precision. Moreover, there was no significant difference in sensitivity between multiplex and singleplex assays, ensuring reliable detection across various plant tissues. This rapid, sensitive, and specific diagnostic assay will provide a valuable tool at ports of entry to prevent the entry of GBNV into the United States.
| Item Type: | Article |
|---|---|
| Divisions: | Global Research Program - Accelerated Crop Improvement |
| CRP: | UNSPECIFIED |
| Uncontrolled Keywords: | groundnut bud necrosis orthotospovirus (GBNV), Orthotospovirus arachinecrosis, phytosanitary measures, TaqMan RT-qPCR, quarantine pathogen |
| Subjects: | Mandate crops > Groundnut Others > Plant Virology |
| Depositing User: | Mr Nagaraju T |
| Date Deposited: | 02 Jun 2025 04:24 |
| Last Modified: | 02 Jun 2025 04:24 |
| URI: | http://oar.icrisat.org/id/eprint/13088 |
| Official URL: | https://pmc.ncbi.nlm.nih.gov/articles/PMC12031075/ |
| Projects: | UNSPECIFIED |
| Funders: | United States Department of Agriculture’s Animal and Plant Health Inspection Service (USDA-APHIS), the Georgia Peanut Commission, United States Department of Agriculture (USDA) |
| Acknowledgement: | The authors acknowledge R.K. Jain at IARI, New Delhi, India, for sharing the GBNV isolate. S.B. acknowledges the team members of the UGA virology team for their support in manuscript preparation. We also acknowledge the anonymous reviewers for their time and providing critical feedback improving the manuscript. |
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