One-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) for detection of tomato torrado virus

Budziszewska, Marta; Wieczorek, Przemysław; Obrępalska‐Stęplowska, Aleksandra

doi:10.1007/s00705-016-2774-2

Cited by 24 publications

(17 citation statements)

References 31 publications

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“…However, the detection limit of the conventional PCR (20 pg RNA). These results are in line with those of previous studies 20 – 24 . Interestingly, the current assay is more sensitive than the previously reported one 16 .…”

Section: Discussionsupporting

confidence: 94%

Detection of Novel duck reovirus (NDRV) using visual reverse transcription loop-mediated isothermal amplification (RT-LAMP)

Cai

Liang

et al. 2018

Sci Rep

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Here we present a visual reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for detecting the gene encoding the σB major outer-capsid protein of novel duck reovirus (NDRV). A set of primers, composed of two outer primers, two inner primers and two loop primers, was designed based on the gene of interest. The LAMP reaction was conducted in a traditional laboratory water bath at 65 °C for 50 min. We compared the performance of calcein/Mn2+ and SYBR Green I dyes, as well as electrophoresis on agarose gel stained with GoldView nucleic acid dye to detect the RT-LAMP-amplified products and all assays could be employed to discriminate between positive and negative specimens in visible or UV light. Our data showed that there is no cross-reaction with other viruses and the RT-LAMP technique displayed high sensitivity for detecting NDRV with a minimal detection limit of 200 fg RNA input. This assay was more sensitive than conventional PCR in detecting NDRV both in natural and experimental infection. In conclusion, the RT-LAMP technique was remarkably sensitive, specific, rapid, simple and profitable for the identification of NDRV.

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Section: Discussionsupporting

confidence: 94%

Detection of Novel duck reovirus (NDRV) using visual reverse transcription loop-mediated isothermal amplification (RT-LAMP)

Cai

Liang

et al. 2018

Sci Rep

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“…Generally, four to six primers are designed that recognize several regions of the target sequence, which gives high specificity in LAMP assay. Recently, LAMP assay been applied to the rapid detection of tomato torrado virus in tomato [2]. Several workers found that LAMP shows more sensitive than quantitative real-time PCR assays [19,21,25].…”

Section: Introductionmentioning

confidence: 99%

Rapid detection of tomato leaf curl Bengaluru virus through loop mediated isothermal amplification assay

2017

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A loop-mediated isothermal amplification (LAMP) technique was employed to develop a simple and rapid method for the detection of tomato leaf curl Bangalore virus (ToLCBaV) in diseased plants of tomato (). Six sets of primers were designed for LAMP technique targeting the conserved AC1 region and successfully detected ToLCBaV. No reaction was detected in the tissues of healthy plants by either the LAMP or the polymerase chain reaction (PCR). The LAMP products can be visualized by presence or absence of turbidity and staining (0.2 μL for 25 μL LAMP product) directly in the tube with nucleic acid stain dye which allowed easy detection. Sensitivity of LAMP assay is 100 times of conventional PCR technique. Although, both the LAMP and the PCR methods were capable of detecting ToLCBaV in infected tissues of tomato, the LAMP method would be more useful than the PCR method for detection of ToLCBaV infection in tomato plants because it is more rapid, simple and accurate method.

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“…These primers hybridize with the stem-loops, with an exception of the loops hybridized by the inner primers and prime strand displacement DNA synthesis [24]. Currently, six primers are widely used in many diagnostic protocols to allow better specificity and sensitivity [30][31][32].…”

Section: Lamp Accelerationmentioning

confidence: 99%

Loop Mediated Isothermal Amplification: Principles and Applications in Plant Virology

Panno

Matić

Tiberini³

et al. 2020

Plants

156

111

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In the last decades, the evolution of molecular diagnosis methods has generated different advanced tools, like loop-mediated isothermal amplification (LAMP). Currently, it is a well-established technique, applied in different fields, such as the medicine, agriculture, and food industries, owing to its simplicity, specificity, rapidity, and low-cost efforts. LAMP is a nucleic acid amplification under isothermal conditions, which is highly compatible with point-of-care (POC) analysis and has the potential to improve the diagnosis in plant protection. The great advantages of LAMP have led to several upgrades in order to implement the technique. In this review, the authors provide an overview reporting in detail the different LAMP steps, focusing on designing and main characteristics of the primer set, different methods of result visualization, evolution and different application fields, reporting in detail LAMP application in plant virology, and the main advantages of the use of this technique.

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One-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) for detection of tomato torrado virus

Cited by 24 publications

References 31 publications

Detection of Novel duck reovirus (NDRV) using visual reverse transcription loop-mediated isothermal amplification (RT-LAMP)

Detection of Novel duck reovirus (NDRV) using visual reverse transcription loop-mediated isothermal amplification (RT-LAMP)

Rapid detection of tomato leaf curl Bengaluru virus through loop mediated isothermal amplification assay

Loop Mediated Isothermal Amplification: Principles and Applications in Plant Virology

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