Rapid and specific detection of Yam mosaic virus by reverse-transcription recombinase polymerase amplification
Yam mosaic virus (YMV; genus Potyvirus ) is considered to cause the most economically important viral disease of yams ( Dioscorea spp.) in West Africa which is the dominant region for yam production globally. Yams are a vegetatively propagated crop and the use of virus-free planting material forms a...
| Main Authors: | , , , , |
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| Format: | Journal Article |
| Language: | Inglés |
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Elsevier
2015
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| Online Access: | https://hdl.handle.net/10568/74423 |
| _version_ | 1855517380779704320 |
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| author | Silva, G. Bömer, M. Nkere, C. Kumar, P. Lava Seal, S.E. |
| author_browse | Bömer, M. Kumar, P. Lava Nkere, C. Seal, S.E. Silva, G. |
| author_facet | Silva, G. Bömer, M. Nkere, C. Kumar, P. Lava Seal, S.E. |
| author_sort | Silva, G. |
| collection | Repository of Agricultural Research Outputs (CGSpace) |
| description | Yam mosaic virus (YMV; genus Potyvirus ) is considered to cause the most economically important viral disease of yams ( Dioscorea spp.) in West Africa which is the dominant region for yam production globally. Yams are a vegetatively propagated crop and the use of virus-free planting material forms an essential component of disease control. Current serological and PCR-based diagnostic methods for YMV are time consuming involving a succession of target detection steps. In this study, a novel assay for specific YMV detection is described that is based on isothermal reverse transcription-recombinase polymerase ampli- fication (RT-exoRPA). This test has been shown to be reproducible and able to detect as little as 14 pg/ l of purified RNA obtained from an YMV-infected plant, a sensitivity equivalent to that obtained with the reverse transcription-polymerase chain reaction (RT-PCR) in current general use. The RT-exoRPA assay has, however, several advantages over the RT-PCR; positive samples can be detected in less than 30 min, and amplification only requires a single incubation temperature (optimum 37 ? C). These features make the RT-exoRPA assay a promising candidate for adapting into a field test format to be used by yam breeding programmes or certification laboratoriesYam mosaic virus (YMV; genus Potyvirus ) is considered to cause the most economically important viral disease of yams ( Dioscorea spp.) in West Africa which is the dominant region for yam production globally. Yams are a vegetatively propagated crop and the use of virus-free planting material forms an essential component of disease control. Current serological and PCR-based diagnostic methods for YMV are time consuming involving a succession of target detection steps. In this study, a novel assay for specific YMV detection is described that is based on isothermal reverse transcription-recombinase polymerase ampli- fication (RT-exoRPA). This test has been shown to be reproducible and able to detect as little as 14 pg/ l of purified RNA obtained from an YMV-infected plant, a sensitivity equivalent to that obtained with the reverse transcription-polymerase chain reaction (RT-PCR) in current general use. The RT-exoRPA assay has, however, several advantages over the RT-PCR; positive samples can be detected in less than 30 min, and amplification only requires a single incubation temperature (optimum 37 ? C). These features make the RT-exoRPA assay a promising candidate for adapting into a field test format to be used by yam breeding programmes or certification laboratories |
| format | Journal Article |
| id | CGSpace74423 |
| institution | CGIAR Consortium |
| language | Inglés |
| publishDate | 2015 |
| publishDateRange | 2015 |
| publishDateSort | 2015 |
| publisher | Elsevier |
| publisherStr | Elsevier |
| record_format | dspace |
| spelling | CGSpace744232024-08-29T11:41:25Z Rapid and specific detection of Yam mosaic virus by reverse-transcription recombinase polymerase amplification Silva, G. Bömer, M. Nkere, C. Kumar, P. Lava Seal, S.E. yams dioscorea mosaic virus diagnosis virology Yam mosaic virus (YMV; genus Potyvirus ) is considered to cause the most economically important viral disease of yams ( Dioscorea spp.) in West Africa which is the dominant region for yam production globally. Yams are a vegetatively propagated crop and the use of virus-free planting material forms an essential component of disease control. Current serological and PCR-based diagnostic methods for YMV are time consuming involving a succession of target detection steps. In this study, a novel assay for specific YMV detection is described that is based on isothermal reverse transcription-recombinase polymerase ampli- fication (RT-exoRPA). This test has been shown to be reproducible and able to detect as little as 14 pg/ l of purified RNA obtained from an YMV-infected plant, a sensitivity equivalent to that obtained with the reverse transcription-polymerase chain reaction (RT-PCR) in current general use. The RT-exoRPA assay has, however, several advantages over the RT-PCR; positive samples can be detected in less than 30 min, and amplification only requires a single incubation temperature (optimum 37 ? C). These features make the RT-exoRPA assay a promising candidate for adapting into a field test format to be used by yam breeding programmes or certification laboratoriesYam mosaic virus (YMV; genus Potyvirus ) is considered to cause the most economically important viral disease of yams ( Dioscorea spp.) in West Africa which is the dominant region for yam production globally. Yams are a vegetatively propagated crop and the use of virus-free planting material forms an essential component of disease control. Current serological and PCR-based diagnostic methods for YMV are time consuming involving a succession of target detection steps. In this study, a novel assay for specific YMV detection is described that is based on isothermal reverse transcription-recombinase polymerase ampli- fication (RT-exoRPA). This test has been shown to be reproducible and able to detect as little as 14 pg/ l of purified RNA obtained from an YMV-infected plant, a sensitivity equivalent to that obtained with the reverse transcription-polymerase chain reaction (RT-PCR) in current general use. The RT-exoRPA assay has, however, several advantages over the RT-PCR; positive samples can be detected in less than 30 min, and amplification only requires a single incubation temperature (optimum 37 ? C). These features make the RT-exoRPA assay a promising candidate for adapting into a field test format to be used by yam breeding programmes or certification laboratories 2015-09 2016-05-25T11:59:07Z 2016-05-25T11:59:07Z Journal Article https://hdl.handle.net/10568/74423 en Limited Access Elsevier Silva, G., Bömer, M., Nkere, C., Kumar, P.L., & Seal, S.E. (2015). Rapid and specific detection of yam mosaic virus by reverse-transcription recombinase polymerase amplification. Journal of Virological Methods, 222, 138-144. |
| spellingShingle | yams dioscorea mosaic virus diagnosis virology Silva, G. Bömer, M. Nkere, C. Kumar, P. Lava Seal, S.E. Rapid and specific detection of Yam mosaic virus by reverse-transcription recombinase polymerase amplification |
| title | Rapid and specific detection of Yam mosaic virus by reverse-transcription recombinase polymerase amplification |
| title_full | Rapid and specific detection of Yam mosaic virus by reverse-transcription recombinase polymerase amplification |
| title_fullStr | Rapid and specific detection of Yam mosaic virus by reverse-transcription recombinase polymerase amplification |
| title_full_unstemmed | Rapid and specific detection of Yam mosaic virus by reverse-transcription recombinase polymerase amplification |
| title_short | Rapid and specific detection of Yam mosaic virus by reverse-transcription recombinase polymerase amplification |
| title_sort | rapid and specific detection of yam mosaic virus by reverse transcription recombinase polymerase amplification |
| topic | yams dioscorea mosaic virus diagnosis virology |
| url | https://hdl.handle.net/10568/74423 |
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