Development of a RT-LAMP assay for real-time detection of criniviruses infecting tomato

Mayu Kobayashi; Takaaki Mashiko; Fariha Wilisiani; Sedyo Hartono; Hisashi Nishigawa; Tomohide Natsuaki; Yutaro Neriya

doi:10.1016/j.jviromet.2022.114662

Development of a RT-LAMP assay for real-time detection of criniviruses infecting tomato

J Virol Methods. 2023 Feb:312:114662. doi: 10.1016/j.jviromet.2022.114662. Epub 2022 Nov 28.

Authors

Mayu Kobayashi¹, Takaaki Mashiko², Fariha Wilisiani³, Sedyo Hartono⁴, Hisashi Nishigawa⁵, Tomohide Natsuaki⁵, Yutaro Neriya⁶

Affiliations

¹ School of Agriculture, Utsunomiya University, Mine-machi 350, Utsunomiya, Tochigi 321-8505, Japan.
² Department of Biological Production Science, Faculty of Agriculture, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-cho, Fuchu, Tokyo 183-8509, Japan.
³ Department of Agrotechnology, Faculty of Agriculture, Institute of Agriculture STIPER, Yogyakarta 55282, Indonesia.
⁴ Virology Laboratory, Faculty of Agriculture, Gadjah Mada University, Bulaksumur, Yogyakarta 55281, Indonesia.
⁵ School of Agriculture, Utsunomiya University, Mine-machi 350, Utsunomiya, Tochigi 321-8505, Japan; Department of Biological Production Science, Faculty of Agriculture, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-cho, Fuchu, Tokyo 183-8509, Japan.
⁶ School of Agriculture, Utsunomiya University, Mine-machi 350, Utsunomiya, Tochigi 321-8505, Japan; Department of Biological Production Science, Faculty of Agriculture, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-cho, Fuchu, Tokyo 183-8509, Japan. Electronic address: neriya@a.utsunomiya-u.ac.jp.

PMID: 36455691
DOI: 10.1016/j.jviromet.2022.114662

Abstract

Yellowing symptoms caused by tomato chlorosis virus (ToCV) and tomato infectious chlorosis virus (TICV), both assigned to the genus Crinivirus, resemble nutrient deficiencies. Therefore, early diagnosis of infections will prevent crop damage and the spread of the viruses. In this study, we established a rapid detection method for ToCV and TICV by reverse transcription-loop-mediated isothermal amplification (RT-LAMP). We first designed primer sets for RT-LAMP specific for ToCV and TICV. Next, by selecting the optimum primer set and determining the optimum conditions for the RT-LAMP reaction, each virus was detected within 50 min by piercing the diseased area of a tomato leaf with a toothpick, immersing the toothpick in the reaction solution, and conducting the RT-LAMP reaction. To verify the accuracy of the procedure, 61 tomato leaf samples showing disease symptoms were collected from five regions of Indonesia, and the RT-LAMP results for the samples were identical to those obtained with the commonly used reverse transcription-polymerase chain reaction.

Keywords: Crinivirus; RT-LAMP; Tomato chlorosis virus; Tomato infectious chlorosis virus.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Crinivirus* / genetics
Plant Diseases
Solanum lycopersicum*

Supplementary concepts

LAMP assay
Tomato chlorosis virus