Development of an on-site plum pox virus detection kit based on immunochromatography

Kensaku Maejima, Misako Himeno, Osamu Netsu, Kazuya Ishikawa, Tetsuya Yoshida, Naoko Fujita, Masayoshi Hashimoto, Ken Komatsu, Yasuyuki Yamaji, Shigetou Namba

Research output: Contribution to journalArticlepeer-review

13 Citations (Scopus)

Abstract

Sharka disease, caused by plum pox virus (PPV), is the most serious viral disease of stone fruit trees. Among the eight known strains of the virus, PPV-D is the most important due to its recent global spread. Although enzyme-linked immunosorbent assay (ELISA) is the most common approach for diagnosing sharka, it involves time-consuming steps and requires expensive equipment and trained technicians. In this study, an on-site PPV detection kit based on immunochromatography was developed using polyclonal antibodies against the coat protein (CP) of a PPV-D isolate. The immunochromatographic (IC) assay kit was as sensitive as a commercial ELISA system for detecting Japanese PPV-D isolates. Moreover, it was easy to use (a one-step procedure), and results could be obtained on-site within 15 min without special laboratory equipment. The IC assay kit detected the virus from every aerial part of symptomatic Japanese apricot trees. In a detailed study of viral localization in leaves, the most suitable plant parts for use in the IC assay were symptomatic mesophyll tissues and the region from the petiole to the main vein. A positive reaction was also observed using the CP of other major (PPV-M and PPV-Rec) and minor (PPV-EA, PPV-W, and PPV-T) strains.

Original languageEnglish
Pages (from-to)176-183
Number of pages8
JournalJournal of General Plant Pathology
Volume80
Issue number2
DOIs
Publication statusPublished - Mar 2014
Externally publishedYes

Keywords

  • Antibody
  • Diagnosis
  • Immunochromatography
  • Japanese apricot
  • Plum pox virus (PPV)
  • Stone fruits

ASJC Scopus subject areas

  • Agronomy and Crop Science
  • Plant Science

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