Development and application of reverse transcription-loop mediated isothermal amplification assay for sensitive detection of groundnut bud necrosis virus infecting potato

被引：3

作者：

Raigond, Baswaraj ^{[1
,2
]}

Pathania, Shruti ^{[1
]}

Verma, Gaurav ^{[1
]}

Bhardwaj, Pooja ^{[1
]}

Kochhar, Tarvinder ^{[1
]}

Chakrabarti, S. K. ^{[1
]}

机构：

[1] ICAR Cent Potato Res Inst, Div Plant Protect, Shimla 171001, Himachal Prades, India

[2] ICAR Indian Inst Millets Res, Ctr Rabi Sorghum, Reg Stn, Solapur 413006, Maharashtra, India

来源：

VIROLOGY | 2023年 / 587卷

关键词：

Groundnut bud necrosis virus; Orthotospovirus arachinecrosis; LAMP; Potato; RT-PCR; Thrips; Detection; RT-LAMP ASSAY; SPOTTED WILT VIRUS; RAPID DETECTION; MOSAIC-VIRUS; TOSPOVIRUS; SUGARCANE; DISEASE; CITRUS; GENE;

D O I：

10.1016/j.virol.2023.109872

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

Reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for detection of groundnut bud necrosis virus (GBNV) causing potato stem necrosis disease. The isothermal temperatures, reaction periods and concentrations of reaction mixture were optimized where, the assay worked well at 65 degrees C for 50 min, 6 U of WarmStart Bst 2.0 DNA polymerase, 1.4 mM dNTPs and 2.0 mM MgSO4. The optimized assay proved to be specific to GBNV with no cross reactivity to other viruses infecting potato in India. The specificity of RT-LAMP assay was found to be 100 fold more sensitive than that of RT-PCR. The developed assay was applied for the detection of GBNV from 80 potato leaf samples where 24 samples were found infected which was confirmed by RT-PCR. It was concluded that the RT-LAMP assay developed for detection of GBNV was specific, sensitive and suitable for its use in virus indexing under potato seed production programme.

引用

页数：7

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