Rapid and Specific Detection of Apple stem grooving virus by Reverse Transcription-recombinase Polymerase Amplification

Kim, Nam‐Yeon; Jeong-hak, Oh，; Lee, Su-Heon; Kim, Hongsup; Moon, Jae Sun; Jeong, Rae‐Dong

doi:10.5423/ppj.nt.06.2018.0108

Cited by 27 publications

(8 citation statements)

References 13 publications

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“…Compared with traditional molecular detection technologies, the LF-RPA assay is faster and more convenient. However, false positive signals sometimes appear in the negative control (Aebischer et al 2014 ; Kim et al 2018 ). Therefore, the steps of the LF-RPA process require further optimization.…”

Section: Discussionmentioning

confidence: 99%

Rapid and simple detection of Phytophthora cactorum in strawberry using a coupled recombinase polymerase amplification–lateral flow strip assay

Song

et al. 2021

Phytopathol Res

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Phytophthora cactorum is a devastating pathogen that infects a wide range of plants and causes Phytophthora rot disease, which has resulted in great economic losses in crop production. Therefore, the rapid and practicable detection of P. cactorum is important for disease monitoring and forecasting. In this study, we developed a lateral flow recombinase polymerase amplification (LF-RPA) assay for the sensitive visual detection of P. cactorum. Specific primers for P. cactorum were designed based on the ras-related protein gene Ypt1; all 10 P. cactorum isolates yielded positive detection results, whereas no cross-reaction occurred in related oomycete or fungal species. The detection limit for the LF-RPA assay was 100 fg of genomic DNA under optimized conditions. Combined with a simplified alkaline lysis method for plant DNA extraction, the LF-RPA assay successfully detected P. cactorum in naturally diseased strawberry samples without specialized equipment within 40 min. Thus, the LF-RPA assay developed in this study is a rapid, simple, and accurate method for the detection of P. cactorum, with the potential for further application in resource-limited laboratories.

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Section: Discussionmentioning

confidence: 99%

Rapid and simple detection of Phytophthora cactorum in strawberry using a coupled recombinase polymerase amplification–lateral flow strip assay

Song

et al. 2021

Phytopathol Res

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“…After completing the virus sequencing step, it is possible to develop new sensitive virus detection techniques and replace costly qRT-PCR-based diagnosis testing systems ( Srivastava et al, 2020 ). Some accurate and ultrasensitive nucleic acid-based testing (NAT) tools in development for multiplexed virus infection capable of replacing qRT-PCR are recombinase polymerase amplification (RPA; Kim et al, 2018 ), loop-mediated isothermal amplification (LAMP; Selvaraj et al, 2019 ), and clustered regularly interspaced short palindromic repeats (CRISPR; Kellner et al, 2019 ). Viruses identification by NAT multiplex systems will be fundamental to identifying multiple resistant germplasm, accelerating host molecular marker discovery for resistance traits, and developing resistant varieties.…”

Section: Virus Resistance Breeding and New Molecular Virus Detection ...mentioning

confidence: 99%

The Viral Threat in Cotton: How New and Emerging Technologies Accelerate Virus Identification and Virus Resistance Breeding

Tarazi

Vaslin

2022

Front. Plant Sci.

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Cotton (Gossypium spp. L., Malvaceae) is the world’s largest source of natural fibers. Virus outbreaks are fast and economically devasting regarding cotton. Identifying new viruses is challenging as virus symptoms usually mimic nutrient deficiency, insect damage, and auxin herbicide injury. Traditional viral identification methods are costly and time-consuming. Developing new resistant cotton lines to face viral threats has been slow until the recent use of molecular virology, genomics, new breeding techniques (NBT), remote sensing, and artificial intelligence (AI). This perspective article demonstrates rapid, sensitive, and cheap technologies to identify viral diseases and propose their use for virus resistance breeding.

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“…Thus, a simpler detection method that also exhibits high specificity, sensitivity, and reliability is urgently needed. Although a recombinase polymerase amplification method was recently reported as a more rapid (~40 min) detection method for ASGV in pear, the method was also reported to frequently yield non-specific amplification bands, as a result of low reaction temperatures (37−42°C) (Kim et al, 2018).…”

Section: Introductionmentioning

confidence: 99%

A Reliable Reverse Transcription Loop-Mediated Isothermal Amplification Assay for Detecting Apple stem grooving virus in Pear

Lee

Jeong

2022

Res. Plant Dis

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Apple stem grooving virus (ASGV) is a high-risk viral pathogen that infects many types of fruit trees, especially pear and apple, and causes serious economic losses across the globe. Thus, rapid and reliable detection assay is needed to identify ASGV infection and prevent its spread. A reliable reverse transcription loop-mediated isothermal amplification (RT-LAMP) was developed, optimize, and evaluated for the coding region of coat protein of ASGV in pear leaf. The developed RT-LAMP facilitated the simple screening of ASGV using visible fluorescence and electrophoresis. The optimized reaction conditions for the RT-LAMP were 63°C for 50 min, and the results showed high specificity and 100-fold greater sensitivity than the reverse transcription polymerase chain reaction. In addition, the reliability of the RT-LAMP was validated using field-collected pear leaves. Furthermore, the potential application of paper-based RNA isolation, combined with RT-LAMP, was also evaluated for detecting ASGV from field-collected samples. These assays could be widely applied to ASGV detection in field conditions and to virus-free certification programs.

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Rapid and Specific Detection of Apple stem grooving virus by Reverse Transcription-recombinase Polymerase Amplification

Cited by 27 publications

References 13 publications

Rapid and simple detection of Phytophthora cactorum in strawberry using a coupled recombinase polymerase amplification–lateral flow strip assay

Rapid and simple detection of Phytophthora cactorum in strawberry using a coupled recombinase polymerase amplification–lateral flow strip assay

The Viral Threat in Cotton: How New and Emerging Technologies Accelerate Virus Identification and Virus Resistance Breeding

A Reliable Reverse Transcription Loop-Mediated Isothermal Amplification Assay for Detecting Apple stem grooving virus in Pear

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