Rapid detection of Prunus necrotic ringspot virus using magnetic nanoparticle-assisted reverse transcription loop-mediated isothermal amplification

Zong, Xiaojuan; Wang, Wenwen; Wei, Hairong; Wang, Jiawei; Chen, Xin; Xu, Li; Zhu, Dong; Tan, Yue

doi:10.1016/j.jviromet.2014.07.033

Cited by 19 publications

(10 citation statements)

References 28 publications

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“…2). Also, LAMP sensitivity for detection of PYMPV, ToLCSiV or TYMoV was similar to those reported for begomoviruses TYLCV and ToLCBaV and other plant viruses such as Potato virus Y (PVY), Prunus necrotic ringspot virus (PNRSV), Tomato torrado virus (ToTV) and Southern tomato virus (STV) (Fukuta et al 2003;Almasi et al 2013;Zong et al 2014;Przewodowska et al 2015;Budziszewska et al 2016;Elvira-González et al 2017;Arutselvan et al 2017).…”

Section: Author´s Reply: All Changes Have Been Donesupporting

confidence: 68%

Fast detection by loop-mediated isothermal amplification (LAMP) of the three begomovirus species infecting tomato in Panama

et al. 2017

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Potato yellow mosaic Panama virus (PYMPV), Tomato leaf curl Sinaloa virus (ToLCSiV) and Tomato yellow mottle virus (TYMoV) of genus Begomovirus (family Geminiviridae) are the only three begomovirus species detected infecting tomato (Solanum lycopersicum L.) in Panama. PYMPV, ToLCSiV and TYMoV induce symptoms of stunting, yellowing, curling, distortion of leaves and reduction of fruit size and cause important economic loses. A loop-mediated amplification under isothermal conditions (LAMP) assay was developed for the individual detection of these three begomovirus species by using a set of three primer pairs specific per each one of them. Amplification products were visualized by gel electrophoresis or direct Gel-Red staining of DNA into the reaction tube. PYMPV, ToLCSiV and TYMoV were detected in total DNA extracts obtained from different plant tissues such as leaves, stems, flowers, fruits and roots of infected tomato plants collected in different production regions of Panama. LAMP sensitivity was similar to that of conventional PCR but, the first procedure was faster and cheaper than the last one. Moreover, all three viruses were successfully detected by LAMP and not by conventional PCR from sap extracts

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Section: Author´s Reply: All Changes Have Been Donesupporting

confidence: 68%

Fast detection by loop-mediated isothermal amplification (LAMP) of the three begomovirus species infecting tomato in Panama

et al. 2017

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“…Moreover, if used with thermostable reverse transcriptase, this method can be applied to detect pathogens with an RNA genome [ 19 , 21 ]. To date, RT-LAMP has been used to detect several plant viruses, including members of the genera Potyvirus [ 21 , 22 ], Comovirus [ 23 ], Ilarvirus [ 24 ], and Crinivirus [ 25 ].…”

Section: Introductionmentioning

confidence: 99%

One-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) for detection of tomato torrado virus

Budziszewska

Wieczorek

Obrępalska‐Stęplowska

2016

Arch Virol

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‘Torrado’ disease caused by tomato torrado virus (ToTV) is responsible for considerable losses in tomato production. Therefore, a one-step reverse transcription loop-mediated isothermal amplification protocol for early and fast detection of ToTV isolates has been developed. The RNA extracted from ToTV-infected plants was tested using this protocol with a set of six primers specific for the Vp35 coat protein gene sequence. The amplified products were analyzed using amplification curves, electrophoresis, and direct staining of DNA. The sensitivity of the protocol was tenfold higher than that of conventional RT-PCR. This new protocol is inexpensive, rapid, simple, and very sensitive.

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“…Field samples tested by using RT-CPA-NATSC demonstrated a 100% agreement with RT-PCR and DAS-ELISA. In X Zong ’s research, the detectability of RT-LAMP is 19.6 × 2 −6 ng/μL total RNA and 19.6 × 2 −4 ng/μL total RNA of RT-PCR whereas the limit of RT-CPA-NATSC is 48 × 10 −4 ng/μL total RNA, which indicate RT-CPA-NATSC is a high sensitivity and specificity of this method on the PNRSV detection 14 .…”

Section: Discussionmentioning

confidence: 93%

“…The result shows only two of seventeen samples tested positive by ELISA, whereas RT-PCR showed all samples but one produced a virus-specific band 13 . While RT-PCR and RT-LAMP are sensitive enough to detect PNRSV in dormant branch compared with ELISA 14 , 15 , the former requires running gels which is time-consuming as well as increase the risk of contamination, and the latter has relative low specificity due to the use of staining dyes for color development of the amplicon 16 . Real-time fluorescence RT-PCR is a rapid detection method for PNRSV but requires the expensive equipment and reagents 17 .…”

Section: Introductionmentioning

confidence: 99%

Rapid Detection of Prunus Necrotic Ringspot Virus by Reverse Transcription-cross-priming Amplification Coupled with Nucleic Acid Test Strip Cassette

Huo

Qiu

et al. 2017

Sci Rep

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Prunus necrotic ringspot virus (PNRSV) is one of the most devastating viruses to Prunus spp. In this study, we developed a diagnostic system RT-CPA-NATSC, wherein reverse transcription-cross-priming amplification (RT-CPA) is coupled with nucleic acid test strip cassette (NATSC), a vertical flow (VF) visualization, for PNRSV detection. The RT-CPA-NATSC assay targets the encoding gene of the PNRSV coat protein with a limit of detection of 72 copies per reaction and no cross-reaction with the known Prunus pathogenic viruses and viroids, demonstrating high sensitivity and specificity. The reaction is performed on 60 °C and can be completed less than 90 min with the prepared template RNA. Field sample test confirmed the reliability of RT-CPA-NATSC, indicating the potential application of this simple and rapid detection method in routine test of PNRSV.

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Rapid detection of Prunus necrotic ringspot virus using magnetic nanoparticle-assisted reverse transcription loop-mediated isothermal amplification

Cited by 19 publications

References 28 publications

Fast detection by loop-mediated isothermal amplification (LAMP) of the three begomovirus species infecting tomato in Panama

Fast detection by loop-mediated isothermal amplification (LAMP) of the three begomovirus species infecting tomato in Panama

One-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) for detection of tomato torrado virus

Rapid Detection of Prunus Necrotic Ringspot Virus by Reverse Transcription-cross-priming Amplification Coupled with Nucleic Acid Test Strip Cassette

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