Multiplex PCR for the detection of African cassava mosaic virus and East African cassava mosaic Cameroon virus in cassava

A multiplex PCR was developed for simultaneous detection of African cassava mosaic virus (ACMV) and East African cassava mosaic Cameroon virus (EACMCV) in cassava affected with cassava mosaic disease (CMD). One set of three primers consisting of an upstream primer common for both viruses and two dow...

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Autores principales: Alabi, O.J., Kumar, P. Lava, Naidu, R.A.
Formato: Journal Article
Lenguaje:Inglés
Publicado: Elsevier 2008
Materias:
Acceso en línea:https://hdl.handle.net/10568/90862
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author Alabi, O.J.
Kumar, P. Lava
Naidu, R.A.
author_browse Alabi, O.J.
Kumar, P. Lava
Naidu, R.A.
author_facet Alabi, O.J.
Kumar, P. Lava
Naidu, R.A.
author_sort Alabi, O.J.
collection Repository of Agricultural Research Outputs (CGSpace)
description A multiplex PCR was developed for simultaneous detection of African cassava mosaic virus (ACMV) and East African cassava mosaic Cameroon virus (EACMCV) in cassava affected with cassava mosaic disease (CMD). One set of three primers consisting of an upstream primer common for both viruses and two down stream virus-specific primers were designed for simultaneous amplification of 368 base pair (bp) and 650 bp DNA fragments specific to the replicase gene of ACMV and EACMCV, respectively. Similarly, a second set of three primerswere designed for simultaneous amplification of 540 bp and 655 bp fragments specific to the coat protein gene of EACMCV and ACMV, respectively. Primers that can amplify a 171 bp fragment of the large subunit of ribulose bisphosphate carboxylase oxygenase Lwere included as an internal control in these assays to determine the reliability of multiplex PCR. A simplified, cost-effective and rapid sample preparation method was adapted in place of the conventional plant DNA extraction procedure for multiplex PCR detection of ACMV and EACMCV. The method was validated using CMD-infected cassava samples obtained from farmers’ fields in Nigeria. The multiplex PCR is useful for reliable assessment of the prevalence of CMBs in epidemiological studies and for crop improvement and phytosanitary programs in African countries.
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spelling CGSpace908622024-08-27T10:35:09Z Multiplex PCR for the detection of African cassava mosaic virus and East African cassava mosaic Cameroon virus in cassava Alabi, O.J. Kumar, P. Lava Naidu, R.A. multiplex pcr begomovirus african cassava mosaic virus sub saharan africa whitefly east african cassava mosaic cameroon virus virology A multiplex PCR was developed for simultaneous detection of African cassava mosaic virus (ACMV) and East African cassava mosaic Cameroon virus (EACMCV) in cassava affected with cassava mosaic disease (CMD). One set of three primers consisting of an upstream primer common for both viruses and two down stream virus-specific primers were designed for simultaneous amplification of 368 base pair (bp) and 650 bp DNA fragments specific to the replicase gene of ACMV and EACMCV, respectively. Similarly, a second set of three primerswere designed for simultaneous amplification of 540 bp and 655 bp fragments specific to the coat protein gene of EACMCV and ACMV, respectively. Primers that can amplify a 171 bp fragment of the large subunit of ribulose bisphosphate carboxylase oxygenase Lwere included as an internal control in these assays to determine the reliability of multiplex PCR. A simplified, cost-effective and rapid sample preparation method was adapted in place of the conventional plant DNA extraction procedure for multiplex PCR detection of ACMV and EACMCV. The method was validated using CMD-infected cassava samples obtained from farmers’ fields in Nigeria. The multiplex PCR is useful for reliable assessment of the prevalence of CMBs in epidemiological studies and for crop improvement and phytosanitary programs in African countries. 2008-12 2018-02-06T12:15:09Z 2018-02-06T12:15:09Z Journal Article https://hdl.handle.net/10568/90862 en Limited Access Elsevier Alabi, O.J., Kumar, P.L. & Naidu, R.A. (2008). Multiplex PCR for the detection of African cassava mosaic virus and East African cassava mosaic Cameroon virus in cassava. Journal of Virological Methods, 154(1), 111-120.
spellingShingle multiplex pcr
begomovirus
african cassava mosaic virus
sub saharan africa
whitefly
east african cassava mosaic cameroon virus
virology
Alabi, O.J.
Kumar, P. Lava
Naidu, R.A.
Multiplex PCR for the detection of African cassava mosaic virus and East African cassava mosaic Cameroon virus in cassava
title Multiplex PCR for the detection of African cassava mosaic virus and East African cassava mosaic Cameroon virus in cassava
title_full Multiplex PCR for the detection of African cassava mosaic virus and East African cassava mosaic Cameroon virus in cassava
title_fullStr Multiplex PCR for the detection of African cassava mosaic virus and East African cassava mosaic Cameroon virus in cassava
title_full_unstemmed Multiplex PCR for the detection of African cassava mosaic virus and East African cassava mosaic Cameroon virus in cassava
title_short Multiplex PCR for the detection of African cassava mosaic virus and East African cassava mosaic Cameroon virus in cassava
title_sort multiplex pcr for the detection of african cassava mosaic virus and east african cassava mosaic cameroon virus in cassava
topic multiplex pcr
begomovirus
african cassava mosaic virus
sub saharan africa
whitefly
east african cassava mosaic cameroon virus
virology
url https://hdl.handle.net/10568/90862
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