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

被引：4

作者：

Kobayashi, Mayu ^{[1
,5
]}

Mashiko, Takaaki ^{[2
,6
]}

Wilisiani, Fariha ^{[3
]}

Hartono, Sedyo ^{[4
]}

Nishigawa, Hisashi ^{[1
,2
]}

Natsuaki, Tomohide ^{[1
,2
]}

Neriya, Yutaro ^{[1
,2
]}

机构：

[1] Utsunomiya Univ, Sch Agr, Mine machi 350, Utsunomiya, Tochigi 3218505, Japan

[2] Tokyo Univ Agr & Technol, Fac Agr, Dept Biol Prod Sci, 3-5-8 Saiwai cho, Fuchu, Tokyo 1838509, Japan

[3] Inst Agr STIPER, Fac Agr, Dept Agrotechnol, Yogyakarta 55282, Indonesia

[4] Gadjah Mada Univ, Fac Agr, Virol Lab, Yogyakarta 55281, Indonesia

[5] Tobu Agr Off, Agr Promot Div, 60-27 Nishihoncho, Ota, Gunma 3730033, Japan

[6] Takii & Co Ltd, Plant Breeding & Expt Stn, 1360 Hari, Konan, Shiga 5203231, Japan

来源：

JOURNAL OF VIROLOGICAL METHODS | 2023年 / 312卷

关键词：

Tomato chlorosis virus; Tomato infectious chlorosis virus; Crinivirus; RT-LAMP; ISOTHERMAL AMPLIFICATION ASSAY; YAM MOSAIC-VIRUS; CHLOROSIS VIRUS; SAMPLES; RNA;

D O I：

10.1016/j.jviromet.2022.114662

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页数：7

共 24 条

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