Link to home

A One-Step Reverse Transcription-Polymerase Chain Reaction System for the Simultaneous Detection and Identification of Multiple Tospovirus Infections

February 2005 , Volume 95 , Number  2
Pages  166 - 171

Hiroyuki Uga and Shinya Tsuda

First author: Saitama Prefectural Agriculture and Forestry Research Center, Kuki, Saitama 346-0037, Japan; and second author: Department of Plant Pathology, National Agricultural Research Center, Tsukuba 305-8666, Japan

Go to article:
Accepted for publication 21 October 2004.

A one-step reverse transcription-polymerase chain reaction (RT-PCR) method has been developed for the simultaneous detection and identification of multiple tospoviruses that infect plants. The RT-PCR system is composed of six primers in a single tube: a universal degenerate primer and five virus species-specific primers. Amplifications resulted in an 848-bp PCR product for Watermelon silver mottle virus, 709-bp for Tomato spotted wilt virus, 589-bp for Impatiens necrotic spot virus, 511-bp for Melon yellow spot virus, and a 459-bp amplicon for Iris yellow spot virus. This system enables the simultaneous detection of at least three types of tospovirus infections, in addition to their species identities, from five possible tospoviruses studied, on the basis of their S RNA combinations. This multiplex RT-PCR system was applied to the detection of tospovirus in ornamental crops cultivated in fields and shows potential for epidemiological studies.

© 2005 The American Phytopathological Society