High-throughput multiplex real-time PCR assay for the simultaneous quantification of DNA and RNA viruses infecting cassava plants
Aims: To develop a multiplex TaqMan-based real-time PCR assay (qPCR) for the simultaneous detection and quantification of both RNA and DNA viruses affecting cassava (Manihot esculenta) in eastern Africa. Methods and Results: The diagnostic assay was developed for two RNA viruses; Cassava brown stre...
| Autores principales: | , , , , , , |
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| Formato: | Journal Article |
| Lenguaje: | Inglés |
| Publicado: |
Oxford University Press
2016
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| Materias: | |
| Acceso en línea: | https://hdl.handle.net/10568/76987 |
| _version_ | 1855540057268551680 |
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| author | Otti, G. Bouvaine, S. Kimata, B. Mkamillo, G. Kumar, P. Lava Tomlins, Keith I. Maruthi, M.N. |
| author_browse | Bouvaine, S. Kimata, B. Kumar, P. Lava Maruthi, M.N. Mkamillo, G. Otti, G. Tomlins, Keith I. |
| author_facet | Otti, G. Bouvaine, S. Kimata, B. Mkamillo, G. Kumar, P. Lava Tomlins, Keith I. Maruthi, M.N. |
| author_sort | Otti, G. |
| collection | Repository of Agricultural Research Outputs (CGSpace) |
| description | Aims: To develop a multiplex TaqMan-based real-time PCR assay (qPCR) for the simultaneous detection and quantification of both RNA and DNA viruses affecting cassava (Manihot esculenta) in eastern Africa.
Methods and Results: The diagnostic assay was developed for two RNA viruses; Cassava brown streak virus (CBSV) and Uganda cassava brown streak virus (UCBSV) and two predominant DNA viruses; African cassava mosaic virus (ACMV) and East African cassava mosaic virus (EACMV), which cause the economically important cassava brown streak disease (CBSD) and cassava mosaic disease (CMD) respectively. Our method, developed by analysing PCR products of viruses, was highly sensitive to detect target viruses from very low quantities of 4–10 femtograms. Multiplexing did not diminish sensitivity or
accuracy compared to uniplex alternatives. The assay reliably detected and quantified four cassava viruses in field samples where CBSV and UCBSV synergy was observed in majority of mixed-infected varieties.
Conclusions: We have developed a high-throughput qPCR diagnostic assay capable of specific and sensitive quantification of predominant DNA and RNA viruses of cassava in eastern Africa.
Significance and Impact of the Study: The qPCR methods are a great improvement on the existing methods and can be used for monitoring virus spread as well as for accurate evaluation of the cassava varieties for virus resistance. |
| format | Journal Article |
| id | CGSpace76987 |
| institution | CGIAR Consortium |
| language | Inglés |
| publishDate | 2016 |
| publishDateRange | 2016 |
| publishDateSort | 2016 |
| publisher | Oxford University Press |
| publisherStr | Oxford University Press |
| record_format | dspace |
| spelling | CGSpace769872023-12-08T19:36:04Z High-throughput multiplex real-time PCR assay for the simultaneous quantification of DNA and RNA viruses infecting cassava plants Otti, G. Bouvaine, S. Kimata, B. Mkamillo, G. Kumar, P. Lava Tomlins, Keith I. Maruthi, M.N. cassava dna plant diseases rna pcr Aims: To develop a multiplex TaqMan-based real-time PCR assay (qPCR) for the simultaneous detection and quantification of both RNA and DNA viruses affecting cassava (Manihot esculenta) in eastern Africa. Methods and Results: The diagnostic assay was developed for two RNA viruses; Cassava brown streak virus (CBSV) and Uganda cassava brown streak virus (UCBSV) and two predominant DNA viruses; African cassava mosaic virus (ACMV) and East African cassava mosaic virus (EACMV), which cause the economically important cassava brown streak disease (CBSD) and cassava mosaic disease (CMD) respectively. Our method, developed by analysing PCR products of viruses, was highly sensitive to detect target viruses from very low quantities of 4–10 femtograms. Multiplexing did not diminish sensitivity or accuracy compared to uniplex alternatives. The assay reliably detected and quantified four cassava viruses in field samples where CBSV and UCBSV synergy was observed in majority of mixed-infected varieties. Conclusions: We have developed a high-throughput qPCR diagnostic assay capable of specific and sensitive quantification of predominant DNA and RNA viruses of cassava in eastern Africa. Significance and Impact of the Study: The qPCR methods are a great improvement on the existing methods and can be used for monitoring virus spread as well as for accurate evaluation of the cassava varieties for virus resistance. 2016-05 2016-09-09T12:18:00Z 2016-09-09T12:18:00Z Journal Article https://hdl.handle.net/10568/76987 en Limited Access Oxford University Press Otti, G., Bouvaine, S., Kimata, B., Mkamillo, G., Kumar, P., Tomlins, K. & Maruthi, M.N. (2016). High throughput multiplex real time PCR assay for the simultaneous quantification of DNA and RNA viruses infecting cassava plants. Journal of applied microbiology 120(5), pp.1346-1356. |
| spellingShingle | cassava dna plant diseases rna pcr Otti, G. Bouvaine, S. Kimata, B. Mkamillo, G. Kumar, P. Lava Tomlins, Keith I. Maruthi, M.N. High-throughput multiplex real-time PCR assay for the simultaneous quantification of DNA and RNA viruses infecting cassava plants |
| title | High-throughput multiplex real-time PCR assay for the simultaneous quantification of DNA and RNA viruses infecting cassava plants |
| title_full | High-throughput multiplex real-time PCR assay for the simultaneous quantification of DNA and RNA viruses infecting cassava plants |
| title_fullStr | High-throughput multiplex real-time PCR assay for the simultaneous quantification of DNA and RNA viruses infecting cassava plants |
| title_full_unstemmed | High-throughput multiplex real-time PCR assay for the simultaneous quantification of DNA and RNA viruses infecting cassava plants |
| title_short | High-throughput multiplex real-time PCR assay for the simultaneous quantification of DNA and RNA viruses infecting cassava plants |
| title_sort | high throughput multiplex real time pcr assay for the simultaneous quantification of dna and rna viruses infecting cassava plants |
| topic | cassava dna plant diseases rna pcr |
| url | https://hdl.handle.net/10568/76987 |
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