Abstract
' Candidatus Liberibacter asiaticus' (Las) is the most prevalent bacterium associated with huanglongbing, which is one of the most destructive diseases of citrus. In this paper, an extremely rapid and simple method for field detection of Las from leaf samples, based on recombinase polymerase amplification (RPA), is described. Three RPA primer pairs were designed and evaluated. RPA amplification was optimized so that it could be accomplished within 10 min. In combination with DNA crude extraction by a 50-fold dilution after 1 min of grinding in 0.5 M sodium hydroxide and visual detection via fluorescent DNA dye (positive samples display obvious green fluorescence while negative samples remain colorless), the whole detection process can be accomplished within 15 min. The sensitivity and specificity of this RPA-based method were evaluated and were proven to be equal to those of real-time PCR. The reliability of this method was also verified by analyzing field samples.
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Claims 6
Claim · model-written summary
The optimized RPA assay could amplify within 10 minutes.
- Interactor
- ‘Candidatus Liberibacter Asiaticus’
Candidatus Liberibacter asiaticust0pathogen - Outcome
- RPA amplificatione0
- Value
- 10 mine0
CitrusOrganisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Citrus Huanglongbing Associated with ‘Candidatus Liberibacter Asiaticus’”
e0“RPA amplification was optimized so that it could be accomplished within 10 min.”
Claim · model-written summary
The full RPA detection process could be completed within 15 minutes.
- Interactor
- ‘Candidatus Liberibacter Asiaticus’Candidatus Liberibacter asiaticust0pathogen
- Outcome
- detection processe0
- Value
- 15 mine0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Citrus Huanglongbing Associated with ‘Candidatus Liberibacter Asiaticus’”
e0“the whole detection process can be accomplished within 15 min”
Claim · model-written summary
Positive samples showed green fluorescence whereas negative samples were colorless.
- Interactor
- ‘Candidatus Liberibacter Asiaticus’Candidatus Liberibacter asiaticust0pathogen
- Outcome
- obvious green fluorescence while negative samples remain colorlesse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Citrus Huanglongbing Associated with ‘Candidatus Liberibacter Asiaticus’”
e0“positive samples display obvious green fluorescence while negative samples remain colorless”
Claim · model-written summary
RPA method sensitivity was equal to real-time PCR.
- Interactor
- ‘Candidatus Liberibacter Asiaticus’Candidatus Liberibacter asiaticust0pathogen
- Outcome
- sensitivitye0
- Direction
- equal toe0
- Comparator
- real-time PCRe0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Citrus Huanglongbing Associated with ‘Candidatus Liberibacter Asiaticus’”
e0“The sensitivity and specificity of this RPA-based method were evaluated and were proven to be equal to those of real-time PCR.”
Claim · model-written summary
RPA method specificity was equal to real-time PCR.
- Interactor
- ‘Candidatus Liberibacter Asiaticus’Candidatus Liberibacter asiaticust0pathogen
- Outcome
- specificitye0
- Direction
- equal toe0
- Comparator
- real-time PCRe0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Citrus Huanglongbing Associated with ‘Candidatus Liberibacter Asiaticus’”
e0“The sensitivity and specificity of this RPA-based method were evaluated and were proven to be equal to those of real-time PCR.”
Claim · model-written summary
Analysis of field samples verified the reliability of the RPA method.
- Interactor
- ‘Candidatus Liberibacter Asiaticus’Candidatus Liberibacter asiaticust0pathogen
- Outcome
- reliabilitye0
- Setting
- field samplese0
Not reportedComparatorValue
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Citrus Huanglongbing Associated with ‘Candidatus Liberibacter Asiaticus’”
e0“The reliability of this method was also verified by analyzing field samples.”