ISHS Acta Horticulturae 386: XVI International Symposium on Fruit Tree Virus diseases
DETECTION OF APPLE PROLIFERATION AND OTHER MLOS BY IMMUNO-CAPTURE PCR (IC-PCR) |
| Authors: | J. Rajan, M.F. Clark |
| Abstract:
We have developed a sensitive immunocapture-PCR detection method in which the target MLO is selectively captured by specific antibody adsorbed on thermostable microtitre plates, following which MLO DNA is released and amplified using either specific or universal primers. The method has been applied successfully to several MLOs in Catharanthus roseus, including the MLOs associated with European aster yellows, peach yellow leafroll, Molieres disease and apple proliferation. Differentiation of these MLOs was achieved using either polyclonal or monoclonal antibodies as the capture agent. We have also successfully detected and identified apple proliferation MLO in apple bark using rabbit polyclonal antibodies to capture the MLO, followed by amplification with universal primers. All samples from infected trees gave a product which was characteristic of apple proliferation MLO by RFLP analysis. No product was obtained from healthy trees or from infected bark samples added to microplate wells coated with antibodies against other MLOs. | |
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