Reverse transcription loop-mediated isothermal amplification based rapid detection of Sugarcane mosaic virus and Sugarcane streak mosaic virus associated with mosaic disease of sugarcane

Anandakumar, L.; Bagyalakshmi, K.; Muthuramalingam, T. Raja; Nithya, K.; Parameswari, B.; Viswanathan, R.

doi:10.1007/s42360-020-00219-w

Cited by 10 publications

(6 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…These products can be visually detected using agarose gel electrophoresis or through colour changes visualised by adding fluorescent dyes to the reaction mixture (Notomi et al, 2000;Parida, Sannarangaiah, Dash, & Rao, 2008;Waliullah et al, 2020). Currently, the RT-LAMP technique has been successfully used in the detection of many RNA plant pathogens (Anandakumar et al, 2020;Bhat et al, 2013;He, Xue, Xu, & Wang, 2016;Liu et al, 2010;Zhang et al, 2020). The colorimetric LAMP method has also been successfully used under field conditions making it an attractive choice where laboratory and technical resources are limited (Keizerweerd, Chandra, & Grisham, 2015;He et al, 2016;Zhao et al, 2018).…”

mentioning

confidence: 99%

Rapid visual detection of Japanese hornwort mosaic virus infecting Angelica sinensis by reverse transcription loop‐mediated isothermal amplification

Zhang

Wang

Xie

et al. 2020

Annals of Applied Biology

View full text Add to dashboard Cite

Japanese hornwort mosaic virus (JHMV; genus Potyvirus, family Potyviridae) is a widespread virus that infects angelica (Angelica sinensis [Oliv.] Diels), an important Chinese herbal medicine plant grown in Gansu, China. JHMV infection has contributed to the deterioration in angelica quality and a reduction in yield. Consequently, there is a need to develop a reliable, simple and rapid detection method to accurately identify JHMV infection and help limit its spread. We describe here, a reverse transcription loop-mediated isothermal amplification (RT-LAMP) developed to detect the coat protein gene of JHMV. RT-LAMP amplification products were assessed through realtime fluorescence detection and by gel electrophoresis and SYBR Green I DNA staining for visual observation. This assay successfully detected JHMV in infected plants without cross reactivity recorded from six other plant viruses. Optimum LAMP reactions were conducted in betaine-free media with 6 mM Mg 2+ at 60 C for 60 min for JHMV. The detection limit was 0.28 pg/ml using RT-LAMP for JHMV plasmids. This detection limit for the RT-LAMP assay was 100 times lower than that of the conventional RT-polymerase chain reaction (RT-PCR) assay. Our field survey of angelica crops for JHMV using RT-LAMP further demonstrated a higher sensitivity than RT-PCR, detecting 78% versus 72%. Agreement (κ) between the results obtained from the two tests was 0.844. We found RT-LAMP is accurate and efficient in diagnosis and potentially improving JHMV disease management and forecasting in A. sinensis in China.

show abstract

mentioning

confidence: 99%

Rapid visual detection of Japanese hornwort mosaic virus infecting Angelica sinensis by reverse transcription loop‐mediated isothermal amplification

Zhang

Wang

Xie

et al. 2020

Annals of Applied Biology

View full text Add to dashboard Cite

show abstract

“…The final pellet was resuspended in 1 mL of 1× PBS and placed on an analytical membrane. The efficiency of the virus particle separation process was confirmed through qRT-PCR, using protocols and gene-specific primers previously developed and exclusively used for ScYLV in our laboratory . The primers used for the qRT-PCR, which are typically employed to detect and quantify various ScYLV-infected sugarcane isolates, were YLS-RT-For-CGTCAAGAGGAACGCCAAGAAAGTC and YLS-RT-Rev-GACGAATTGTCCTGCTAGGCTCGA.…”

Section: Methodsmentioning

confidence: 98%

“…The efficiency of the virus particle separation process was confirmed through qRT-PCR, using protocols and gene-specific primers previously developed and exclusively used for ScYLV in our laboratory. 11 The primers used for the qRT-PCR, which are typically employed to detect and quantify various ScYLV-infected sugarcane isolates, were YLS-RT-For-CGTCAAGAGGAACGCCAA-GAAAGTC and YLS-RT-Rev-GACGAATTGTCCTGCTAGGCTC-GA. The qRT-PCR reactions were performed using Power SYBR Green Master Mix from Applied Biosystems, in two different PCR systems (Veriti and PCR) also from Applied Biosystems.…”

Section: Experimental Design and Diagnosticmentioning

confidence: 99%

Magnetic Nanozyme-Enhanced Rapid ImmunoFlow-Through Assay for the Femtomolar Detection of Sugarcane Yellow Leaf Virus

Thangavelu,

PK,

Ramasamy

et al. 2024

ACS Agric. Sci. Technol.

Self Cite

View full text Add to dashboard Cite

Plant diseases significantly threaten global food security, with numerous historical instances of devastating epidemics. This risk is particularly acute in key agricultural and food crops, such as sugarcane. Although recent advancements in molecular diagnostics have improved the detection of sugarcane viruses, these methods are largely confined to lab settings due to their reliance on sophisticated, costly equipment. To overcome this limitation, we have developed a more accessible and cost-effective solution: a magnetic nanozyme-enhanced colorimetric ImmunoFlow-through assay designed for the ultrasensitive detection of sugarcane yellow leaf curl virus (ScYLV). This innovative technique allows for clear optical identification of viral concentrations as low as femtomolar levels. The assay employs cationic magnetic nanoparticles for virus isolation and colorimetric immunolabels for diagnosis, enhancing sensitivity and providing immediate results, comparable to those of established methods like quantitative real-time-polymerase chain reaction (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA). Our assay offers a one-step detection process and a two-step semiquantitative analysis, marking a major breakthrough in plant virus diagnostics. Extensive research into the assay’s design, including its sensing platform, blocking agents, antibody conjugation chemistries, sensitivity, quantification, potential for multiplexing, and field applicability, was carried out. This diagnostic research utilizing Magnetozyme in a flow-through assay represents a pioneering approach to rapid and sensitive diagnosis within plant disease diagnostics. It introduces a promising alternative to traditional molecular diagnostics, potentially transforming plant disease management and enhancing food security globally.

show abstract

“…RT-LAMP is a cost-effective alternative to RT-PCR, as the reaction can be carried out in a heat block or a water bath. It has been shown that RT-LAMP exhibits similar or even higher sensitivity than the conventional RT-PCR (Hasiów-Jaroszewska and Borodynko 2013; Anandakumar et al 2020;Li et al 2022). Taking into account the advantages of the isothermal assay, the main aim of this study was to develop and optimize RT-LAMP with originally designed primers, for rapid detection of genetic variants of MDMV isolates.…”

mentioning

confidence: 99%

“…MDMV and SCMV induce similar symptoms and cause the same viral disease, therefore, specific diagnostic tools are required to distinguish between both viruses. To date, RT-LAMP assay has been successfully optimized for the detection of different viruses infecting ornamental plants (Yao et al 2022), vegetables (Hasiów-Jaroszewska andBorodynko 2013;Budziszewska et al 2016), cereals (Zarzyńska-Nowak et al 2018), sugarcane (Keizer weerd et al 2015Anandakumar et al 2020) and maize (Chen et al 2017;Li et al 2022). Due to genetic differences between MDMV and SCMV, the primer pairs for the RT-LAMP assay were originally designed based on the alignment of CP gene sequences.…”

mentioning

confidence: 99%

Journal of Plant Protection Research

Trzmiel,

Hasiów-Jaroszewska

2022

View full text Add to dashboard Cite

Maize dwarf mosaic virus (MDMV) is a serious and widespread virus pathogen of maize plants. This +ssRNA virus belongs to the Potyvirus genus in the Potyviridae family. Together with sugarcane mosaic virus (SCMV) it causes one of the most important viral diseases on maize crops in the world -maize dwarf mosaic. Both viruses are transmitted in the same non-persistent manner by several aphid species. They induce similar symptoms of leaf mosaic or mottling, stunting and a reduction in plant weight and grain yield. Available MDMV diagnostics include primarily commercialized enzyme-linked immunosorbent assays (ELISA) and reverse transcription-polymerase chain reactions (RT-PCR). Here, laborsaving reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay was optimized for identification of genetically different MDMV isolates. For this purpose, primer sets, MDMVF3/MDMVB3 and MDMVFIP/MDMVBIP amplifying fragments of coat protein coding sequence of MDMV, were used. The specificity of the reaction was verified using three MDMV (-P1, -Sp, -PV0802-DSMZ) and three SCMV (-P1, -PV0368--DSMZ, -PV1207-DSMZ) isolates. Obtained products were visualised by DNA staining, electrophoretic separation as well as by real-time monitoring of the reaction. The sensitivity of RT-LAMP and conventional RT-PCR reactions was comparable. Both methods could detect virus as low as 550 fg • µl -1 of total RNA. This technique has application value for screening MDMV by phytosanitary services.

show abstract

Reverse transcription loop-mediated isothermal amplification based rapid detection of Sugarcane mosaic virus and Sugarcane streak mosaic virus associated with mosaic disease of sugarcane

Cited by 10 publications

References 30 publications

Rapid visual detection of Japanese hornwort mosaic virus infecting Angelica sinensis by reverse transcription loop‐mediated isothermal amplification

Rapid visual detection of Japanese hornwort mosaic virus infecting Angelica sinensis by reverse transcription loop‐mediated isothermal amplification

Magnetic Nanozyme-Enhanced Rapid ImmunoFlow-Through Assay for the Femtomolar Detection of Sugarcane Yellow Leaf Virus

Journal of Plant Protection Research

Contact Info

Product

Resources

About