Phytoparasitica

, 28:141 | Cite as

Potato Y Potyvirus Detection by Immunological and Molecular Techniques in Plants and Aphids

Research Papers
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Abstract

An antiserum against potato Y potyvirus (PVY) was produced and the double antibody sandwich (DAS)-ELISA technique with a detection sensitivity of 10 ng/ml purified virus, was applied. One-step immunocapture (IC) — reverse transcription (RT) — polymerase chain reaction (PCR) was assessed using published primers; a gain in sensitivity of 1000-fold over ELISA was achieved. Furthermore, the use of PCR-ELISA to detect the produced amplicons after their direct adsorption on a microplate offered a supplementary 100-fold gain in sensitivity. Thus, 100 fg/ml purified virus in healthy tobacco extract became readily detectable, as did an infected potato sample at a dilution end point of 10-7, equivalent to 1 ng tissue. This highly sensitive procedure coupled with the print-capture (PC) modification to avoid specific sample preparation, offered a most handy and useful tool for effective PVY detection in single aphids(Myzus persicae).

Key Words

Potato Y potyvirus (PVY) ELISA IC-PCR PCR-ELISA aphids 
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Copyright information

© Springer Science + Business Media B.V. 2000

Authors and Affiliations

  1. 1.Laboratory of VirologyBenaki Phytopathological InstituteKiflssia, AthensGreece

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