Simultaneous detection and typing of plum pox potyvirus (PPV) isolates by heminested-PCR and PCR-ELISA
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AutorOlmos, Antonio; Cambra, Mariano; Dasi, M. A.; Candresse, Thierry; Esteban, Olga; Gorris, María T.; Asensio, M.
Cita bibliográficaOlmos, A., Cambra, M., Dasi, M.A., Candresse, T., Esteban, O., Gorris, M. T., Asensio, M. (1997). Simultaneous detection and typing of plum pox potyvirus (PPV), isolates by heminested-PCR and PCR-ELISA. Journal of virological methods, 68(2), 127-137.
Two techniques for simultaneous detection and typing of plum pox potyvirus (PPV) isolates belonging to the D or M serotypes, heminested PCR (H-PCR) and PCR-ELISA, have been developed. Ten PPV isolates typed using PPV-D and PPV-M specific monoclonal antibodies by ELISA-DASI were used to validate these two methods. The results obtained show a complete coincidence of the nucleic acid-based techniques with the serological data. When serial dilutions of infected plant extracts were assayed, H-PCR and PCR-ELISA were found to be 100 rimes more sensitive than the more conventional immunocapture-PCR (IC-PCR) assay. Testing of 228 PPV-infected fruit tree samples coming from different hosts and locations indicated that so fat-only PPV type D appears to be present in Spain and in Chile. Coupled with print-capture sample preparation (Olmos et al., Nucl. Acids Res. 24, 2192-2193, 1996) the increased sensitivity provided by heminested-PCR allowed the detection of PPV targets of D and M types, in wingless individuals of the aphid vector Aphis gossypii. (C) 1997 Elsevier Science Ireland Ltd.