Development of a real-time RT-PCR SYBR green assay for Tomato ring spot virus in grape

Elwin L. Stewart, Xinshun Qu, Barrie E. Overton, Fred E. Gildow, Nancy G. Wenner, Deborah S. Grove

Research output: Contribution to journalArticle

12 Scopus citations

Abstract

Grapevines infected with Tomato ring spot virus (ToRSV) pose an economic risk for growers in the northeastern United States. This study describes a one-step real-time reverse-transcription polymerase chain reaction (RT-PCR) SYBR Green assay for detecting ToRSV in grapevines. Two newly designed primer pairs based on the ToRSV coat protein gene sequence were evaluated for specificity and optimized for a SYBR Green assay. The primer pair ToRSV1f/1r yielded a 130-bp product with strong primer-dimer products, whereas the primer pair ToRSV2f/2r yielded a 330-bp product with weak primer dimer products. Real-time RT-PCR detected ToRSV in more naturally infected grapevines maintained in the greenhouse than did enzyme-linked immunosorbent assay. The nucleotide sequences of the fragments amplified from grapevine growing in Pennsylvania using real-time PCR were divergent from previously published sequences.

Original languageEnglish (US)
Pages (from-to)1083-1088
Number of pages6
JournalPlant disease
Volume91
Issue number9
DOIs
StatePublished - Sep 1 2007

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All Science Journal Classification (ASJC) codes

  • Agronomy and Crop Science
  • Plant Science

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