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Development of an immunomagnetic capture-reverse transcriptase-PCR assay for three pineapple ampeloviruses
C F Gambley1, A D W Geering, J E Thomas
1Department of Primary Industries and Fisheries, Horticulture and Forestry Science, 80 Meiers Road, Indooroopilly, Queensland 4068, Australia. cherie.gambley@dpi.qld.gov.au
Abstract:
A semi-automated, immunomagnetic capture-reverse transcription PCR (IMC-RT-PCR) assay for the detection of three pineapple-infecting ampeloviruses, Pineapple mealybug wilt-associated virus-1, -2 and -3, is described. The assay was equivalent in sensitivity but more rapid than conventional immunocapture RT-PCR. The assay can be used either as a one- or two-step RT-PCR and allows detection of the viruses separately or together in a triplex assay from fresh, frozen or freeze-dried pineapple leaf tissue. This IMC-RT-PCR assay could be used for high throughput screening of pineapple planting propagules and could easily be modified for the detection of other RNA viruses in a range of plant species, provided suitable antibodies are available.
