Genes Encoding Potential Molecular Mimicry Proteins as the Specific Targets for Detecting Bursaphelenchus xylophilus in PCR and Loop-Mediated Isothermal Amplification Assays

The introduction of the pine wood nematode ( ) to new areas has affected the international forestry industry because this pathogen causes pine wilt disease (PWD). Therefore, methods for the accurate and reliable detection of are essential for controlling and managing this pest. The PCR and Loop-Mediated Isothermal Amplification (LAMP) techniques developed in this study involve species-specific primer sets targeting genes encoding potential molecular mimicry proteins ( , , and ), which are associated with pathogenicity. The PCR and LAMP results revealed that the primers were specific for , , and . Moreover, our LAMP assay targeting conducted at 63°C detected within 20 min and from or within 30 min. The lower limits of detection for the LAMP and PCR assays were 10 pg and 10 ng genomic DNA, respectively, implying these assays may be useful for the rapid detection of in pine forests. Designing primers specific for , , and enabled the relatively rapid detection of isolates as well as or carrying . These primers, which were designed following a thorough functional analysis of key pathogenicity-related genes, may be useful for developing improved assays for the early diagnosis and prevention of PWD.