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Development of a multiplex immunocapture PCR with colourimetric detection for viruses of banana

Sharman, M., Thomas, J. E. and Dietzgen, R. G. (2000) Development of a multiplex immunocapture PCR with colourimetric detection for viruses of banana. Journal of Virological Methods, 89 (1-2). pp. 75-88. ISSN 01660934 (ISSN)

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Article Link(s): https://doi.org/10.1016/S0166-0934(00)00204-4

Abstract

A multiplex, immunocapture PCR (M-IC-PCR) was developed for the simultaneous detection of three viruses from crude sap extracts of banana and plantain (Musa spp.). A reverse transcription step was required for Banana bract mosaic virus and Cucumber mosaic virus, which have ssRNA genomes. The detection of Banana bunchy top virus (ssDNA genome) was not adversely affected by inclusion in this step. All the three viruses could be detected simultaneously from a mixed infection. Identification and detection of individual viruses was achieved through the visualisation of discretely sized PCR amplicons by gel electrophoresis. Alternatively, a colourimetric microplate detection system utilising digoxigenin-labelled virus-specific probes was used. The latter assay was up to five times more sensitive than detection by gel electrophoresis and between 25 and 625 times more sensitive than ELISA for the various viruses. Careful selection of PCR primers was necessary to ensure the detection of a wide range of virus isolates and to avoid detrimental interactions between heterologous templates and primers. (C) 2000 Elsevier Science B.V.

Item Type:Article
Business groups:Horticulture and Forestry Science
Keywords:Abaca Diagnosis Indexing RNA single stranded DNA article controlled study enzyme linked immunosorbent assay fruit nonhuman nucleotide sequence plant virus priority journal reverse transcription polymerase chain reaction virus detection Badnavirus Circoviridae Colorimetry DNA, Viral Enzyme-Linked Immunosorbent Assay Mosaic Viruses Polymerase Chain Reaction RNA, Viral Support, Non-U.S. Gov't Zingiberales
Subjects:Plant culture > Fruit and fruit culture > Culture of individual fruits or types of fruit > Bananas
Plant pests and diseases > Plant pathology
Deposited On:18 Feb 2021 06:52
Last Modified:18 Feb 2021 06:52

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