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Nigeria Agricultural Journal

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Protocol for cost effective detection of cassava mosaic virus

N. Okereke, J. Tomlinson, I. Adams, L. Laurenson, N. Fairless, N. Boonham, A.F. Okunade, J. Onyeka, J. Smith

Abstract


Early detection of cassava mosaic disease (CMD) is an extremely important step in containing the spread of the disease in Africa. Many nucleic acid based detection tools have been developed for CMD diagnosis but although these methods are specific and sensitive for their target DNA, they are not fast, cost effective, can‘t be used in poorly resourced laboratories and are not portable enough to be used in field settings. This study makes use of crude, but fast method of DNA extraction, alkaline  polyethylene glycol, and an equally fast Loop mediated Isothermal amplification method of detection that doesn‘t require purified DNA. As a diagnostic method for CMD detection, these protocols take approximately 1 hour to conclude, compared to a normal polymerase chain reaction protocol that can take up to 24 hours to conclude.


Keywords: Polyethylene glycol method, Loop mediated Isothermal Amplification, and Polymerase Chain Reaction




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