Development of a reverse-transcription loop-mediated isothermal amplification assay for the detection of Tobacco mild green mosaic virus (TMGMV)
•An RT-LAMP assay for TMGMV detection was developed and reported for the first time.•The primer set designed for the assay was highly specific for TMGMV in the RT-LAMP assay.•The RT-LAMP assay was more convenient and economical, and 10 times more sensitive than the corresponding RT-PCR assay for TMG...
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Veröffentlicht in: | Journal of virological methods 2021-12, Vol.298, p.114277-114277, Article 114277 |
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Sprache: | eng |
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Zusammenfassung: | •An RT-LAMP assay for TMGMV detection was developed and reported for the first time.•The primer set designed for the assay was highly specific for TMGMV in the RT-LAMP assay.•The RT-LAMP assay was more convenient and economical, and 10 times more sensitive than the corresponding RT-PCR assay for TMGMV detection, and was applied successfully for virus testing of pepper field samples and seed.
Tobacco mild green mosaic virus (TMGMV), a member species of the genus Tobamovirus, infects pepper (Capsicum annuum) and a number of other economically important species in the Solanaceae family. TMGMV infections had seriously impacted pepper production worldwide, including China. A reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed to detect TMGMV in pepper field samples and seed. This assay was based on four primers that matched to six sequences in the C-terminal region of the TMGMV genome. RT-LAMP assay could detect the presence of the virus in 3.0 × 10−7 µg of total RNA extract from pepper leaves, which was ten times more sensitive than the corresponding reverse-transcription polymerase chain reaction (RT-PCR) assay. This method specifically detected TMGMV but not the closely related species of the same genus Pepper mild mottle virus, Cucumber green mottle mosaic virus and Tomato mosaic virus. In addition, the use of SYBR Green I facilitated the detection of the TMGMV RT-LAMP products by the naked eye. These results indicated that the RT-LAMP assay was a simple, sensitive, specific and affordable diagnostic tool that has the potential to detect and monitor TMGMV infection in field samples. |
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ISSN: | 0166-0934 1879-0984 |
DOI: | 10.1016/j.jviromet.2021.114277 |