Abstract
Grapevine fleck virus (GFkV) is a graft-transmissible pathogen that threatens grapevine production in rootstocks and hybrid cultivars. Its early and accurate detection is essential for certifying virus-free planting materials. This study developed and validated a reverse-transcription droplet digital polymerase chain reaction (RT-ddPCR) assay for the specific and absolute quantification of GFkV in grapevine tissues. The assay demonstrated high specificity, showing no cross-reactivity with other major grapevine viruses. Meanwhile, RT-ddPCR exhibited approximately 10-fold higher sensitivity than RT-quantitative PCR (RT-qPCR), with a detection limit of 0.73 copies/μl. Both RT-ddPCR and RT-qPCR exhibited high linearity (R2=0.9997 and 0.9983, respectively). In field validation, GFkV was detected in 84.8% (56/66) of the samples via RT-ddPCR, whereas RT-qPCR detected the virus in 53.0% (35/66) of the samples. These results indicate that RT-ddPCR is a highly sensitive and reliable method for GFkV detection in grapevines.
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Claims 7
Claim · model-written summary
The RT-ddPCR assay showed high specificity without cross-reactivity with other grapevine viruses.
- Interactor
- grapevine virusese0pathogen
- Outcome
- high specificitye0
- Comparator
- other major grapevine virusese0
VitisOrganisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“in Grapevine”
e0“The assay demonstrated high specificity, showing no cross-reactivity with other major grapevine viruses.”
Claim · model-written summary
RT-ddPCR sensitivity was approximately tenfold higher than RT-qPCR.
- Outcome
- sensitivitye0
- Direction
- highere0
- Value
- approximately 10-folde0
- Comparator
- RT-quantitative PCR (RT-qPCR)e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“in Grapevine”
e0“RT-ddPCR exhibited approximately 10-fold higher sensitivity than RT-quantitative PCR (RT-qPCR), with a detection limit of 0.73 copies/μl.”
Claim · model-written summary
RT-ddPCR had a detection limit of 0.73 copies/μl.
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“in Grapevine”
e0“RT-ddPCR exhibited approximately 10-fold higher sensitivity than RT-quantitative PCR (RT-qPCR), with a detection limit of 0.73 copies/μl.”
Claim · model-written summary
RT-ddPCR had high linearity, R2=0.9997.
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“in Grapevine”
e0“Both RT-ddPCR and RT-qPCR exhibited high linearity (R2=0.9997 and 0.9983, respectively).”
Claim · model-written summary
RT-qPCR had high linearity, R2=0.9983.
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“in Grapevine”
e0“Both RT-ddPCR and RT-qPCR exhibited high linearity (R2=0.9997 and 0.9983, respectively).”
Claim · model-written summary
RT-ddPCR detected GFkV in 84.8% of field samples.
- Interactor
- GFkVGrapevine fleck viruse0pathogen
- Outcome
- GFkV was detectede0
- Value
- 84.8% (56/66)e0
- Comparator
- RT-qPCRe0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“in Grapevine”
e0“In field validation, GFkV was detected in 84.8% (56/66) of the samples via RT-ddPCR, whereas RT-qPCR detected the virus in 53.0% (35/66) of the samples.”
Claim · model-written summary
RT-qPCR detected GFkV in 53.0% of field samples.
- Interactor
- GFkVGrapevine fleck viruse0pathogen
- Outcome
- detected the viruse0
- Value
- 53.0% (35/66)e0
- Comparator
- RT-ddPCRe0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“in Grapevine”
e0“In field validation, GFkV was detected in 84.8% (56/66) of the samples via RT-ddPCR, whereas RT-qPCR detected the virus in 53.0% (35/66) of the samples.”