Simple and Rapid Detection of Potato leafroll virus by Reverse Transcription Loop-mediated Isothermal Amplification

Ju, Ho-Jong

doi:10.5423/ppj.2011.27.4.385

Cited by 25 publications

(10 citation statements)

References 29 publications

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“…Colorimetric (Goto et al 2009) and fluorescent (Tomita et al 2008) variants of the assay were also developed. RT-LAMP has been used to detect various pathogens, including plant viruses such as Plum pox virus (Varga and James 2006), Potato leafroll virus (Ahmadi et al 2013;Ju 2011), Bean pod mottle virus (Wei et al 2012), Cassava brown streak virus (Tomlinson et al 2013), Pepino mosaic virus (Hasiów-Jaroszewska and Borodynko 2013), Papaya leaf distortion virus (Shen et al 2014) and Cucurbit chlorotic yellows virus (Wang et al 2014). The agarose gel based and turbidimetric variants of the method were also adopted for detection of PVY (Nie 2005;Zhao et al 2012).…”

Section: Introductionmentioning

confidence: 99%

A One-Step, Real-Time Reverse Transcription Loopmediated Isothermal Amplification Assay to Detect Potato Virus Y

et al. 2015

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Potato viruses such as Potato virus Y (PVY) cause diseases that affect potato quality and thus damage potato production worldwide. Current tests for viral infection use double-antibody sandwich enzyme linked immunosorbent assays (DAS ELISA) or reverse transcription polymerase chain reaction (RT-PCR)/real-time RT-PCR. Despite many advantages, these assays have a number of drawbacks that affect cost and time of diagnosis. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) allows fast detection of target RNA. Here, we developed a closed-tube real-time RT LAMP assay for fluorescent detection of PVY. Specific RT-LAMP primers were designed to target the conserved region of the sequence encoding the PVY coat protein. The assay was specific and facilitated sensitive PVY detection in a single tube at 65°C. The time-topositive values depended on the PVY concentration in tested samples. The effectiveness of RT LAMP in testing field-grown plants compared favorably with DAS ELISA and RT-PCR; under the tested conditions, RT-LAMP was about 1000-fold more sensitive than DAS ELISA and lateral flow assay (LFA) and about 10-fold more sensitive than RT PCR. Thus, this fluorescent RT-LAMP assay has great potential for routine detection of PVY. 1000 veces más sensible que DAS ELISA y que el ensayo de flujo lateral, y como 10 veces más sensible que RT-PCR. La efectividad de RT-LAMP al probarla en plantas en el campo se comparó favorablemente con DAS ELISA y RT-PCR. En consecuencia, presenta un gran potencial para detección de rutina del PVY.

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Section: Introductionmentioning

confidence: 99%

A One-Step, Real-Time Reverse Transcription Loopmediated Isothermal Amplification Assay to Detect Potato Virus Y

et al. 2015

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“…Compared to other techniques of PLRV detection, the proposed silver-enhanced LFIA is more sensitive than previously-described LFIAs in conventional format with an LOD of 30 ng/mL (Kondakova, Butenko, Skurat, & Drygin, 2016) and more rapid than tissue print immunoassay with a duration of 4-5 h (Samsatly, Jawhari, Najjar, Sobh, & Abou-Jawdah, 2014). Moreover, due to possibility of on-site application, this LFIA is a good alternative for PLRV assays based on reverse transcription polymerase chain reactions (Cating, Funke, Kaur, Hamm, & Frost, 2015;Ju, 2011;Zhang et al, 2017) or loop-mediated isothermal amplification (Ahmadi, Almasi, Fatehi, Struik, & Moradi, 2013;Almasi, Manesh, Jafary, & Dehabadi, 2013).…”

Section: Comparison Of Lfias Without Enhancement and With Silver Enhamentioning

confidence: 97%

Silver-enhanced lateral flow immunoassay for highly-sensitive detection of potato leafroll virus

Panferov

Safenkova

Byzova

et al. 2017

Food and Agricultural Immunology

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Rapid non-laboratory screening of plants for pathogenic viruses crucially influences crop yields in modern agricultural technologies. The aim of this study was to develop a highly-sensitive lateral flow immunoassay (LFIA) for rapid detection of potato leafroll virus (PLRV), an infectious agent of one of the most widespread potato diseases. The proposed LFIA combines the formation of sandwich immune complexes with gold nanoparticles (GNP) as labels and silver enhancement. The enhancement stage was realized using mixture of silver lactate and hydroquinone and subsequent addition of chloride-containing buffer to stop silver reduction. LFIA with silver enhancement was 15 times more sensitive (detection limit 0.2 ng/mL; 15 min) compared with conventional LFIA (detection limit 3 ng/mL; 10 min). The enhanced LFIA detected PLRV in leaves' extracts of infected potato in dilutions higher than enzyme-linked immunosorbent assay.

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“…The fluorescently labeled amplified product can be quantified using a fluorescence reader, which is light weight, weather-proof, battery operated, GPS enabled, and offers rapid, on-farm end-point detection. Subsequently, RT-LAMP method has been developed for detection of Potato leaf roll virus [9], Pepino mosaic virus [10], Tomato torrado virus [11], and Southern tomato virus [12]. Warghane et al [13] showed the sensitivity of RT-LAMP is 100 times more than RT-PCR using the RNA extracted from the CTV infected leaf samples.…”

Section: Introductionmentioning

confidence: 99%

A rapid detection tool for VT isolates of Citrus tristeza virus by immunocapture-reverse transcriptase loop-mediated isothermal amplification assay

Selvaraj

Maheshwari

Hajeri³

et al. 2019

PLoS ONE

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Severe strains of Citrus tristeza virus (CTV) cause quick decline and stem pitting resulting in significant economic losses in citrus production. A immunocapture reverse-transcriptase loop-mediated amplification (IC-RT-LAMP) assay was developed in this study to detect the severe VT strains that are typically associated with severe CTV symptoms. The sensitivity of RT-LAMP assay was determined by tenfold serial dilutions of CA-VT-AT39 RNA, in comparison to one-step RT-droplet digital (dd) PCR. RT-LAMP detected up to 0.002 ng RNA with an amplification time of 10:35 (min:sec.), equivalent to 11.3 copies as determined by one step RT-ddPCR. The RT-LAMP assay specifically detected CA-VT-AT39 RNA and did not cross react with other CTV genotypes tested (T36, T30, RB, S1 and T68). To facilitate rapid on-site detection, the RT-LAMP assay was improved by first capturing the CTV virions from citrus crude leaf sap using CTV-IgG (IC-RT-LAMP), thereby eliminating nucleic acid extraction steps. IC-RT-LAMP assay was optimized with twofold dilutions of CTV-IgG ranging from 1:500 to 1:16,000. The IC-RT-LAMP assay detected the CA-VT-AT39 virions in all dilutions tested. The minimum amplification time was 6:45 (min:sec) with 1:500 and 1:1000 of CTV-IgG dilutions. The limit of detection of IC-RT-LAMP assay with crude leaf sap of CA-VT-AT39 was 1:320 with a maximum amplification time of 9:08 (min:sec). The IC-RT-LAMP assay was validated for VT genotype by comparing to IC-RT-qPCR using the CTV from 40 field tree samples. A 100% agreement was observed between tests, regardless of single or mixed infections of CTV VT with other genotypes. Therefore, the IC-RT-LAMP assay can serve as a useful tool in the management of potentially severe strains of CTV.

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Simple and Rapid Detection of Potato leafroll virus by Reverse Transcription Loop-mediated Isothermal Amplification

Cited by 25 publications

References 29 publications

A One-Step, Real-Time Reverse Transcription Loopmediated Isothermal Amplification Assay to Detect Potato Virus Y

A One-Step, Real-Time Reverse Transcription Loopmediated Isothermal Amplification Assay to Detect Potato Virus Y

Silver-enhanced lateral flow immunoassay for highly-sensitive detection of potato leafroll virus

A rapid detection tool for VT isolates of Citrus tristeza virus by immunocapture-reverse transcriptase loop-mediated isothermal amplification assay

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