Development of a Real-Time PCR Assay to Detect the Kiwifruit Pathogen <i>Pectobacterium actinidiae</i>

<i>Pectobacterium actinidiae</i> is a significant kiwifruit canker pathogen reported from kiwifruit-growing areas in Asia and South America. To improve the diagnostic capability for this bacterium, a probe-based real-time PCR assay was developed for sensitive and fast detection of <i>P. actinidiae</i>. The highly specific assay targets the meta-pathway phenol degradation protein-like locus, unique to <i>P. actinidiae</i>. Validation showed that the assay is highly sensitive, detecting the pathogen down to 0.1 pg of pure genomic DNA or 10 to 50 colony-forming units in plant tissue matrix. To assess the competency of the tested DNA samples and avoid false-negative results, the assay was duplexed with plant internal control. This new method provides a valuable tool for specific and sensitive testing of imported and exported plant material to ensure freedom from <i>P. actinidiae</i> and prevent the spread of this pathogen into new areas.[Formula: see text] Copyright © 2025 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.