Development of one step colorimetric RT-LAMP assays for rapid detection of Apple mosaic virus and Prunus necrotic ringspot virus

Wani, Latief A; Jawa, Priyanka; Khan, J. A.

doi:10.1016/j.jviromet.2023.114729

Cited by 6 publications

(1 citation statement)

References 38 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This finding is consistent with previous studies reported by Wani et al . [ 45 ] and Caruso et al . [ 46 ] for prunus necrotic ringspot virus and TSWV, respectively.…”

Section: Discussionmentioning

confidence: 99%

A reverse transcription loop-mediated isothermal amplification assay for quick detection of tomato mosaic virus

Kirasi,

Ateka,

Avedi

et al. 2024

PLoS ONE

View full text Add to dashboard Cite

Tomato mosaic virus (ToMV), an economically important virus that affects a wide range of crops, is highly contagious, and its transmission is mediated by mechanical means, and through contaminated seeds or planting materials, making its management challenging. To contain its wide distribution, early and accurate detection of infection is required. A survey was conducted between January and May, 2023 in major tomato growing counties in Kenya, namely, Baringo, Kajiado, Kirinyaga and Laikipia, to establish ToMV disease incidence and to collect samples for optimization of the reverse transcription loop-mediated isothermal amplification assay (RT-LAMP) assay. A RT-LAMP assay, utilizing primers targeting the coat protein, was developed and evaluated for its performance. The method was able to detect ToMV in tomato samples within 4:45 minutes, had a 1,000-fold higher sensitivity than conventional reverse transcription polymerase chain reaction (RT-PCR) method and was specific to ToMV. Furthermore, the practical applicability of the assay was assessed using tomato samples and other solanaecous plants. The assay was able to detect the virus in 14 tomato leaf samples collected from the field, compared to 11 samples detected by RT-PCR, further supporting the greater sensitivity of the assay. To make the assay more amenable for on-site ToMV detection, a quick-extraction method based on alkaline polyethylene glycol buffer was evaluated, which permitted the direct detection of the target virus from crude leaf extracts. Due to its high sensitivity, specificity and rapidity, the RT-LAMP method could be valuable for field surveys and quarantine inspections towards a robust management of ToMV infections.

show abstract

“…This finding is consistent with previous studies reported by Wani et al . [ 45 ] and Caruso et al . [ 46 ] for prunus necrotic ringspot virus and TSWV, respectively.…”

Section: Discussionmentioning

confidence: 99%

A reverse transcription loop-mediated isothermal amplification assay for quick detection of tomato mosaic virus

Kirasi,

Ateka,

Avedi

et al. 2024

PLoS ONE

View full text Add to dashboard Cite

show abstract

Development of reverse transcription loop-mediated isothermal amplification (RT-LAMP) for rapid detection of viruses infecting patchouli (Pogostemon cablin)

Bano,

Khan

2024

Arch Microbiol

Self Cite

View full text Add to dashboard Cite

Patchouli (Pogostemon cablin) a highly valued medicinal plant, suffers signi cant economic losses following infection with Broad bean wilt virus 2 (BBWV2) and Peanut stripe virus (PStV). In this study, a eld-based isothermal technique called reverse transcription loop-mediated isothermal ampli cation (RT-LAMP) was established for an early and speci c identi cation of BBWV2 and PStV. The oligo primers for RT-LAMP assay were designed from the conserved domains of the coat protein genes of PStV and BBWV2. The reaction temperature and time duration were optimised to 65 0 C for 60 min, followed by incubation at 85 0 C for 10 min in a water bath. The LAMP amplicons positive for PStV and BBWV2 revealed characteristic ladder-type bands when analysed through gel electrophoresis. Further, a colorimetric assay using a metal ion-based indicator (Hydroxy-napthol Blue, HNB) was conducted to visualize the ampli ed products with the naked eye, thus facilitating accessibility to eld practices. The assays developed in this study were virus-speci c as no cross-ampli cation occurred with other viruses, and were 100 times more sensitive than RT-PCR. Thus, the RT-LAMP assays established in this study are quick, reliable, and cost-effective for the accurate identi cation of BBWV2 and PStV. It will facilitate screening of patchouli planting materials. Thus, may reduce the risk of virus spread and could be helpful in phytosanitary programs.

show abstract

Colorimetric strategies applicable for loop-mediated isothermal amplification

Saifuddin,

Rashid,

Nor Azmi

et al. 2024

Journal of Microbiological Methods

View full text Add to dashboard Cite

Development of one step colorimetric RT-LAMP assays for rapid detection of Apple mosaic virus and Prunus necrotic ringspot virus

Cited by 6 publications

References 38 publications

A reverse transcription loop-mediated isothermal amplification assay for quick detection of tomato mosaic virus

A reverse transcription loop-mediated isothermal amplification assay for quick detection of tomato mosaic virus

Development of reverse transcription loop-mediated isothermal amplification (RT-LAMP) for rapid detection of viruses infecting patchouli (Pogostemon cablin)

Colorimetric strategies applicable for loop-mediated isothermal amplification

Contact Info

Product

Resources

About