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

Elsevier

Available online 28 November 2022, 114662

Journal of Virological MethodsAuthor links open overlay panelHighlights•

RT-LAMP primer sets specific for ToCV and TICV was designed.

The toothpick method allows detection by RT-LAMP without RNA extraction.

This is the first report of RT-LAMP detection of TICV.

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

Tomato chlorosis virus

Tomato infectious chlorosis virus

Crinivirus

RT-LAMP

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