Development of loop-mediated isothermal amplification assay for rapid detection of genetically different wheat dwarf virus isolates

Trzmiel, Katarzyna; Hasiów‐Jaroszewska, Beata

doi:10.1007/s11033-020-05846-0

Cited by 9 publications

(5 citation statements)

References 16 publications

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“…RT-LAMP assays have been described for several plant viruses with RNA genomes [ 26 ], including wheat viruses [ 32 ], but not for WSSMV. LAMP-based protocols for wheat virus detection described to date mainly entail the extraction of DNA [ 33 , 34 ] or RNA [ 35 ] or reverse transcription into cDNA [ 36 ], few of them being directly applicable to crude plant extracts [ 27 , 37 ]. This study developed a protocol of RT-LAMP for the direct detection of WSSMV in 30 minutes from crude durum wheat leaf extracts.…”

Section: Discussionmentioning

confidence: 99%

Rapid and specific detection of wheat spindle streak mosaic virus using RT-LAMP in durum wheat crude leaf extract

Marra,

Mussano,

Pinton

et al. 2024

PLoS ONE

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To accurately determine the spread of any pathogen, including plant viruses, a quick, sensitive, cost-effective, point-of-care diagnostic assay is necessary. Wheat spindle streak mosaic virus (WSSMV) is a Bymovirus, transmitted by the plasmodiophorid Polymyxa graminis Led, which causes yellow mosaic and reduces the grain yield in wheat. Currently, detection protocols for WSSMV use ELISA or more sensitive PCR-based approaches requiring specialized laboratory and personnel. A protocol for reverse transcription loop mediated isothermal amplification (RT-LAMP) has been developed and optimized for the rapid detection of viruses using crude extracts from wheat leaves. The protocol was specific for WSSMV detection, while no reaction was observed with SBCMV or SBWMV, the non-target viruses transmitted by the same vector. The RT-LAMP assay was shown to be as sensitive as the one-step WSSMV specific RT-PCR. The RT-LAMP assay can be performed under field conditions using a portable instrument, and can help the actual spread of WSSMV, an aspect of this virus not yet well understood, to be explored.

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Section: Discussionmentioning

confidence: 99%

Rapid and specific detection of wheat spindle streak mosaic virus using RT-LAMP in durum wheat crude leaf extract

Marra,

Mussano,

Pinton

et al. 2024

PLoS ONE

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“…Lee et al [ 40 ] described a procedure to detect wheat streak mosaic virus (WSMV) during quarantine inspections of imported cereals, where LAMP is performed on cDNA following RNA extraction. A LAMP protocol has been proposed also for a DNA virus (i.e., wheat dwarf virus) [ 41 ].…”

Section: Discussionmentioning

confidence: 99%

Fast and Sensitive Detection of Soil-Borne Cereal Mosaic Virus in Leaf Crude Extract of Durum Wheat

Marra

D’Errico

Montemurro

et al. 2022

Viruses

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Soil-borne cereal mosaic virus (SBCMV) is a furovirus with rigid rod-shaped particles containing an ssRNA genome, transmitted by Polymyxa graminis Led., a plasmodiophorid that can persist in soil for up to 20 years. SBCMV was reported on common and durum wheat and it can cause yield losses of up to 70%. Detection protocols currently available are costly and time-consuming (real-time PCR) or have limited sensitivity (ELISA). To facilitate an efficient investigation of the real dispersal of SBCMV, it is necessary to develop a new detection tool with the following characteristics: no extraction steps, very fast results, and high sensitivity to allow pooling of a large number of samples. In the present work, we have developed a reverse transcription loop-mediated isothermal amplification (RT-LAMP) protocol with such characteristics, and we have compared it with real-time PCR. Our results show that the sensitivity of LAMP and real-time PCR on cDNA and RT-LAMP on crude extracts are comparable, with the obvious advantage that RT-LAMP produces results in minutes rather than hours. This paves the way for extensive field surveys, leading to a better knowledge of the impact of this virus on wheat health and yield.

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“…The detection of potato viruses typically requires field samples to be sent to a well-equipped laboratory for nucleic acid extraction and purification followed by RT-PCR and melt curve analysis. More recently, there has been an effort to replace the complex RT-PCR step with simpler isothermal amplification methods [ 38 , 39 , 40 , 41 , 42 ]. Therefore, we set out to develop a rapid, simple, and inexpensive approach for the detection of potato viruses that (a) minimizes sample processing; (b) provides reliable samples for the rapid and robust on-site diagnosis of infections by isothermal amplification; and (c) allows on-site verification by LFD analysis.…”

Section: Discussionmentioning

confidence: 99%

Fast, Precise, and Reliable Multiplex Detection of Potato Viruses by Loop-Mediated Isothermal Amplification

Edgü

Freund

Hartje³

et al. 2020

IJMS

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Potato is an important staple food crop in both developed and developing countries. However, potato plants are susceptible to several economically important viruses that reduce yields by up to 50% and affect tuber quality. One of the major threats is corky ringspot, which is a tuber necrosis caused by tobacco rattle virus (TRV). The appearance of corky ringspot symptoms on tubers prior to commercialization results in ≈ 45% of the tubers being downgraded in quality and value, while ≈ 55% are declared unsaleable. To improve current disease management practices, we have developed simple diagnostic methods for the reliable detection of TRV without RNA purification, involving minimalized sample handling (mini), subsequent improved colorimetric loop-mediated isothermal amplification (LAMP), and final verification by lateral-flow dipstick (LFD) analysis. Having optimized the mini-LAMP-LFD approach for the sensitive and specific detection of TRV, we confirmed the reliability and robustness of this approach by the simultaneous detection of TRV and other harmful viruses in duplex LAMP reactions. Therefore, our new approach offers breeders, producers, and farmers an inexpensive and efficient new platform for disease management in potato breeding and cultivation.

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Development of loop-mediated isothermal amplification assay for rapid detection of genetically different wheat dwarf virus isolates

Cited by 9 publications

References 16 publications

Rapid and specific detection of wheat spindle streak mosaic virus using RT-LAMP in durum wheat crude leaf extract

Rapid and specific detection of wheat spindle streak mosaic virus using RT-LAMP in durum wheat crude leaf extract

Fast and Sensitive Detection of Soil-Borne Cereal Mosaic Virus in Leaf Crude Extract of Durum Wheat

Fast, Precise, and Reliable Multiplex Detection of Potato Viruses by Loop-Mediated Isothermal Amplification

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