Development of the Droplet Digital PCR Method for the Detection and Quantification of Erwinia pyrifoliae

Black shoot blight disease caused by Erwinia pyrifoliae has serious impacts on quality and yield in pear production in Korea; therefore, rapid and accurate methods for its detection are needed. However, traditional detection methods require a great deal of time and fail to achieve absolute quantific...

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Bibliographic Details
Published in:The plant pathology journal 2023-02, Vol.39 (1), p.141-148
Main Authors: He, Lin, Kim, Seong Hwan, Yu, Jun Myoung
Format: Article
Language:English
Subjects:
black shoot blight
droplet digital polymerase chain reaction
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Summary:Black shoot blight disease caused by Erwinia pyrifoliae has serious impacts on quality and yield in pear production in Korea; therefore, rapid and accurate methods for its detection are needed. However, traditional detection methods require a great deal of time and fail to achieve absolute quantification. In the present study, we developed a droplet digital polymerase chain reaction (ddPCR) method for the detection and absolute quantification of E. pyrifoliae using a pair of species-specific primers. The detection range was 103 - 107 copies/ml (DNA templates) and cfu/ml (cell culture templates). This new method exhibited good linearity and repeatability and was validated by absolute quantification of E. pyrifoliae DNA copies from samples of artificially inoculated immature pear fruits. Here, we present the first study of ddPCR assay for the detection and quantification of E. pyrifoliae. This method has potential applications in epidemiology and for the early prediction of black shoot blight outbreaks.
ISSN:1598-2254
2093-9280
DOI:10.5423/PPJ.NT.08.2022.0117