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Simultaneous detection and typing of plum pox potyvirus (PPV) isolates by heminested-PCR and PCR-ELISA

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URI
http://hdl.handle.net/20.500.11939/4259
DOI
10.1016/S0166-0934(97)00120-1
Derechos de acceso
openAccess
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Author
Olmos, Antonio; Cambra, Mariano; Dasi, M. A.; Candresse, Thierry; Esteban, Olga; Gorris, María T.; Asensio, M.
Date
1997
Cita bibliográfica
Olmos, 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.
Abstract
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.
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