Development of an immunomagnetic capture-reverse transcriptase-PCR assay for three pineapple ampeloviruses.
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.
Gambley
C F
CF
Department of Primary Industries and Fisheries, Horticulture and Forestry Science, 80 Meiers Road, Indooroopilly, Queensland 4068, Australia. cherie.gambley@dpi.qld.gov.au
Geering
A D W
AD
Thomas
J E
JE
D031230:Ananas / Q000821:virology*; D029381:Closteroviridae / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification*; D018189:Immunomagnetic Separation / Q000379:methods*; D010935:Plant Diseases / Q000821:virology*; D018515:Plant Leaves / Q000821:virology; D012367:RNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D012367:RNA, Viral