Development of colorimetric and real time loop-mediated isothermal amplification (cr-LAMP) assay for rapid detection of Wheat dwarf virus (WDV)

Çeli̇k, Ali; Morca, Ali Ferhan

doi:10.1016/j.cropro.2021.105786

Cited by 11 publications

(2 citation statements)

References 47 publications

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“…The current study demonstrated that 1.4 mM dNTP concentration was ideal for B. oryzae detection by LAMP assay. The result of the present investigation was found to be parallel with those of [61,74,75].…”

supporting

confidence: 88%

Potential of LAMP assay as a tool for ecological monitoring and on-site detection of Bipolaris oryzae in rice

Logeshwari

Gopalakrishnan

Kamalakannan

et al. 2023

Preprint

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One of the most significant rice seed-borne pathogens, Bipolaris oryzae causes serious yield losses worldwide every year. Traditional seed health testing involves incubation for up to a week, but molecular diagnosis using PCR-based assays requires sophisticated equipment and is not amenable to on-site testing. Loop-mediated isothermal amplification (LAMP) is ideal for field-level diagnosis since it offers quick, high-specific amplification of target template sequences at a single temperature. The exon 2 of the Glycosyl Transferase Family 90 protein of B. oryzae was targeted for designing the LAMP primers. The LAMP assay proposed was incredibly sensitive, detecting the B. oryzaetemplate's presence at concentrations as low as 10 fg at 60 °C in 30 min. The specificity of the test was assessed using genomic DNA retrieved from 22 different phytopathogens. The assay was employed to biologically monitor the existence of this pathogen by testing it on ten weed species in rice fields infected with B. oryzae. The pathogen was detected in 8 weed species except Brachypodium distachyon and Phalaris minor. The assay was used to detect the presence of B. oryzae in seeds from 13 different rice varieties. Seed samples from two cultivars tested negative while the remaining cultivars were confirmed to harbor the pathogen. Using this novel assay, B. oryzae can be detected rapidly in rice seeds and farm soil. The proposed LAMP assay has the potential to be employed in seed health laboratories, plant quarantine stations and on-site detection of B. oryzae in seeds and plants.

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supporting

confidence: 88%

Potential of LAMP assay as a tool for ecological monitoring and on-site detection of Bipolaris oryzae in rice

Logeshwari

Gopalakrishnan

Kamalakannan

et al. 2023

Preprint

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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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Wild Relatives and Their Contributions to Wheat Breeding

Tekin

Emi̇rali̇oğlu²,

Yeken³

et al. 2022

Ancient Wheats

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Development of colorimetric and real time loop-mediated isothermal amplification (cr-LAMP) assay for rapid detection of Wheat dwarf virus (WDV)

Cited by 11 publications

References 47 publications

Potential of LAMP assay as a tool for ecological monitoring and on-site detection of Bipolaris oryzae in rice

Potential of LAMP assay as a tool for ecological monitoring and on-site detection of Bipolaris oryzae in rice

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

Wild Relatives and Their Contributions to Wheat Breeding

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