“Rapid diagnosis of Cucumber mosaic virus in banana plants using a fluorescence-based real-time isothermal reverse transcription-recombinase polymerase amplification assay”

Srivastava, Nishant; Kapoor, Reetika; Kumar, Rakesh; Kumar, Swagat; Saritha, R. K.; Kumar, Sandeep; Baranwal, V. K.

doi:10.1016/j.jviromet.2019.04.024

Cited by 38 publications

(14 citation statements)

References 19 publications

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“…Moreover, the established RPA assay can be completed within 30 min and shows no crossreaction with other selected viruses infecting maize plants. At present, the RPA method is usually combined with LFD to obtain visual results, which is suitable for resource-limited laboratories and field survey . However, RPA products should be diluted prior to LFD detection, which will reduce their sensitivity .…”

Section: Resultsmentioning

confidence: 99%

“…At present, the RPA method is usually combined with LFD to obtain visual results, which is suitable for resource-limited laboratories and field survey. 24 However, RPA products should be diluted prior to LFD detection, which will reduce their sensitivity. 20 Therefore, agarose gel electrophoresis is still suggested as the most common visualization method followed by RPA detection.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

“…RPA can be completed by only one pair of primers to avoid the complicated primer design process. , The RPA assay can be carried out in a temperature range of 37–42 °C in 30 min without any special thermal cycling equipment . RPA can also be improved by combining it with other assays, such as agarose gel electrophoresis, fluorescence-based RT-RPA assay, and RPA-lateral flow dipstick (RPA-LFD) . In particular, agarose gel electrophoresis is the most common method for detection visualization.…”

Section: Introductionmentioning

confidence: 99%

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Recombinase Polymerase Amplification Assay for Simultaneous Detection of Maize Chlorotic Mottle Virus and Sugarcane Mosaic Virus in Maize

et al. 2021

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Maize chlorotic mottle virus (MCMV) can cause maize lethal necrosis (MLN) when coinfected with potyvirids, such as sugarcane mosaic virus (SCMV), maize dwarf mosaic virus, or wheat streak mosaic virus. MLN is often caused by coinfection of MCMV and SCMV, which has been reported in China and several countries of Africa. In this study, a recombinase polymerase amplification (RPA) assay was established for simultaneous detection of MCMV and SCMV in maize. The RPA assay can be completed within 30 min at 38 °C. The primers for the RPA assay were specific since no crossreaction was detected with other selected viruses that infected maize in China. The detection limit of the RPA method was 10 2 copies μL −1 , which was about 10-fold more sensitive than that of the conventional PCR method. Moreover, the RPA assay can be successfully applied to detect maize samples collected in the field. These results demonstrated that the established RPA assay is a rapid and efficient method to conduct simultaneous detection of MCMV and SCMV, which provides an alternative technology for MLN diagnosis.

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Section: Resultsmentioning

confidence: 99%

Section: ■ Results and Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Recombinase Polymerase Amplification Assay for Simultaneous Detection of Maize Chlorotic Mottle Virus and Sugarcane Mosaic Virus in Maize

et al. 2021

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“…The amount of signal produced is directly proportional to the concentration of the amplicon. These approaches were used to develop detection assays for cucumber mosaic virus infecting banana (Srivastava et al ., 2019), cucurbit yellow stunting disorder virus infecting watermelon and squash (Kalischuk et al ., 2020), plum pox virus infecting stone fruits (Zhang et al ., 2014), potato virus Y (Cassedy et al ., 2022), rose rosette virus (Babu et al ., 2017b), yam mosaic virus, and yam mild mosaic virus (Silva et al ., 2018; Table 1).…”

Section: Real‐time Detection Of Rpa Products Based On Fluorescencementioning

confidence: 99%

Onsite detection of plant viruses using isothermal amplification assays

Bhat

Aman

Mahfouz

2022

Plant Biotechnology Journal

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Summary Plant diseases caused by viruses limit crop production and quality, resulting in significant losses. However, options for managing viruses are limited; for example, as systemic obligate parasites, they cannot be killed by chemicals. Sensitive, robust, affordable diagnostic assays are needed to detect the presence of viruses in plant materials such as seeds, vegetative parts, insect vectors, or alternative hosts and then prevent or limit their introduction into the field by destroying infected plant materials or controlling insect hosts. Diagnostics based on biological and physical properties are not very sensitive and are time‐consuming, but assays based on viral proteins and nucleic acids are more specific, sensitive, and rapid. However, most such assays require laboratories with sophisticated equipment and technical skills. By contrast, isothermal‐based assays such as loop‐mediated isothermal amplification (LAMP) and recombinase polymerase amplification (RPA) are simple, easy to perform, reliable, specific, and rapid and do not require specialized equipment or skills. Isothermal amplification assays can be performed using lateral flow devices, making them suitable for onsite detection or testing in the field. To overcome non‐specific amplification and cross‐contamination issues, isothermal amplification assays can be coupled with CRISPR/Cas technology. Indeed, the collateral activity associated with some CRISPR/Cas systems has been successfully harnessed for visual detection of plant viruses. Here, we briefly describe traditional methods for detecting viruses and then examine the various isothermal assays that are being harnessed to detect viruses.

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“…The reactions can be completed in 5-20 min depending on the starting copy number of the targeted template and the size of the amplicon. The RPA assay has been reported in diagnosis of several plant viruses, including those with DNA genomes such as wheat dwarf virus (Glais and Jacquot 2015), banana bunchy top virus (Kapoor et al 2017), citrus yellow mosaic virus (Kumar et al 2018), bean golden yellow mosaic virus, tomato mottle virus, tomato yellow leaf curl virus (Londoño et al 2016;Wang and Yang 2019), milk vetch dwarf virus (Cao et al 2020), and piper yellow mottle virus (Mohandas and Bhat 2020); and with RNAbased genomes such as little cherry virus-2 (Mekuria et al 2014), plum pox virus (Zhang et al 2014;Babujee et al 2019), rose rosette virus (Babu et al 2017), rice blackstreaked dwarf virus (Zhao et al 2019), cucumber mosaic virus (Srivastava et al 2019), maize chlorotic mottle virus (Jiao et al 2019a), apple stem pitting virus (Kim et al 2019), cucumber green mottle mosaic virus (Jiao et al 2019b;Zeng et al 2019), cucurbit yellow stunting disorder virus (Kalischuk et al 2020), and peach latent mosaic viroid (Lee et al 2020). RPA has also been successfully applied for detection of the potyviruses plum pox virus (Zhang et al 2014), yam mosaic virus and yam mild mosaic virus (Silva et al 2015), potato virus Y (Glais and Jacquot 2015;Babujee et al 2019), and chilli veinal mottle virus (Jiao et al 2020).…”

Section: Introductionmentioning

confidence: 99%

Development of a recombinase polymerase amplification combined with lateral flow dipstick assay for rapid and sensitive detection of bean common mosaic virus

et al. 2021

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Bean common mosaic virus (BCMV) is one of the most widespread and damaging viruses of cultivated legumes in the world. In addition to serious yield reduction and germplasm decline, BCMV infection also makes legumes more vulnerable to other pathogens. Early diagnosis of the virus is particularly important in limiting its spread. Recombinase polymerase amplification (RPA) is a novel isothermic amplification technology. The whole reaction can be done outside the laboratory environment after the nucleic acid sample is obtained. In this study, we established a rapid and sensitive RPA combined with the lateral flow dipstick (LFD) assay for detection of BCMV, based on the conserved BCMV coat protein (CP) gene sequence. Specific primers and a probe were designed, which amplify ~ 150 bp CP fragments from BCMV-infected samples under a constant temperature of 37 °C for 20 min. The end-labeled amplification products were detected by high-affinity LFD within 5 min. Sensitivity of this RPA-LFD assay was 1000 times greater than that of the conventional polymerase chain reaction (PCR) assay. Furthermore, when the primers/probe were used against related potyviruses including soybean mosaic virus (SMV), bean yellow mosaic virus (BYMV), and turnip mosaic virus (TuMV), the three potyviruses were not detected, indicating that the assay was BCMV species-specific. The RPA-LFD assay was also successfully applied for the detection of seed-borne BCMV in beans. The RPA-LFD assay has great potential application in the rapid diagnosis of BCMV in the field.

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“Rapid diagnosis of Cucumber mosaic virus in banana plants using a fluorescence-based real-time isothermal reverse transcription-recombinase polymerase amplification assay”

Cited by 38 publications

References 19 publications

Recombinase Polymerase Amplification Assay for Simultaneous Detection of Maize Chlorotic Mottle Virus and Sugarcane Mosaic Virus in Maize

Recombinase Polymerase Amplification Assay for Simultaneous Detection of Maize Chlorotic Mottle Virus and Sugarcane Mosaic Virus in Maize

Onsite detection of plant viruses using isothermal amplification assays

Development of a recombinase polymerase amplification combined with lateral flow dipstick assay for rapid and sensitive detection of bean common mosaic virus

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