Detection of episomal banana streak badnavirus by IC-PCR.
A polymerase chain reaction (PCR) based strategy to detect episomal banana streak badnavirus (BSV) in banana and plantain plants that carry integrated BSV sequences was developed. Antisera used in immuno-capture polymerase chain reaction (IC-PCR) are capable of binding a large number of BSV serotypes. The primers used for PCR are capable of annealing to and amplifying across the aspartic protease-reverse transcriptase domain boundaries of both episomal and integrated BSV sequences and result in similar or identical sequence size fragments from either template. However, we show that under the conditions selected for IC-PCR, nuclear, mitochondrial or chloroplast genomic sequences are not amplified and thus only captured episomal BSV is amplified. IC-PCR is suitable for the large-scale screening of Musa for episomal BSV which is necessary for germplasm movement.
D019195:Badnavirus / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D015139:Blotting, Southern; D018744:DNA, Plant / Q000032:analysis; D005638:Fruit / Q000821:virology*; D007118:Immunoassay / Q000379:methods*; D010935:Plant Diseases / Q000821:virology; D018515:Plant Leaves / Q000821:virology; D010957:Plasmids; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity